Triazolopyridines and triazolopyrimidines that lower stress-induced p-tau

ABSTRACT

In certain embodiments compounds (e.g., triazolopyrimidine(s) and/or triazolopyridine(s)) are provided that are effective to lower p-tau and/or to prevent or reduce elevation of p-tau, particularly in response to stress (e.g., elevated cortisol levels). The 5 compounds are useful in the treatment or prophylaxis of pathologies characterized by abnormal amyloid plaque and or tangle formation and/or deposition.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims benefit of and priority to U.S. Ser. No.62/117,888, filed Feb. 18, 2015, which is incorporated herein byreference in its entirety for all purposes.

STATEMENT OF GOVERNMENTAL SUPPORT

[Not Applicable]

BACKGROUND

Alzheimer's disease (AD) is estimated to afflict more than 20 millionpeople worldwide and is believed to be the most common cause ofdementia. As the World population ages, the number of people withAlzheimer's disease (AD), currently approximately 5.4 million in theUnited States, will continue to rise. Alzheimer's is a neurodegenerativedisease associated with progressive dementia and memory loss. Two keycharacteristics of AD are the accumulation of extracellular depositscontaining aggregated Aβ peptide and neuronal synaptic loss in the AD inspecific brain regions. Although AD pathogenesis is complex, compellinggenetic and biochemical evidence suggest that overproduction of Aβ, orfailure to clear this peptide is the earliest event in the amyloidcascade that lead to AD primarily through amyloid deposition, which ispresumed to be involved in neurofibrillary tangle formation, neuronaldysfunction and microglia activation, that characterize AD-affectedbrain tissues.

Neurofibrillary tangles, along with plaques comprised of Aβ peptide, area pathological hallmark of Alzheimer's Disease (AD).Hyperphosphorylation of the microtubule-stabilizing protein tau leads totangle formation. In people diagnosed with AD, and in our hands usingthe J20 mouse model of AD, the level of tau phosphorylation has theclosest correlation to cognitive impairment. The reversal of taupathology alone can improve memory, even in the presence of high Aβ42 inJ20 mice (Roberson, et al. (2007) Science, 316(5825): 750-754). Evenwhen Aβ plaque load is similar, reduction in tau expression andtherefore tau pathology (tau −) increases performance in the MorrisWater Maze. Tau pathology is “downstream” of Aβ formation in that Aβincreases expression of glycogen synthase kinase 3beta (GSK-3β), anenzyme that can increase phosphorylation of tau. The isoform of Aβ,whether soluble, oligomeric, and/or plaque-bound likely affects GSK-3βexpression, therefore absolute AB level may not be directly related top-tau level. Convergent evidence implicates stress in AD neuropathology(Carroll, et al. (2011) J Neurosci. (40): 14436-14449). Stress exposurecan increase AP production and induce deficits in hippocampal cellproliferation and contextual memory (Wilson et al. (2003) Neurology, 61:1479-1485). Moreover, exposure to a variety of physiological stressorscan activate tau kinases and induce tau phosphorylation (tau-P) inrodents (Dong et al. (2004) Neurosci., 127: 601-609). The corticotropinreleasing factor (CRF) signaling system plays a role in response tostress (Kang et al. (2007) Proc. Natl. Acad. Sci. USA, 104:10673-10678).

SUMMARY

Various embodiments contemplated herein may include, but need not belimited to, one or more of the following:

Embodiment 1

A compound according to the formula:

or a pharmaceutically acceptable salt thereof, wherein: R⁰ is present orabsent, and when present is selected from the group consisting of CHR,NH, O, and NCHR where R is H, alkyl (e.g., a C1-C6 carbon chain), oraryl (e.g., phenyl, substituted phenyl, or heteroaryl); R² is

and R¹ is

or is selected from the group consisting of a substituted orunsubstituted cyclic or heterocycle selected from the group consistingof pyridine, pyrimidine, naphthalene, quinolone, isoquinoline,cinnoline, phenyl, substituted phenyl, oxazole, furan, pyran, isoxazole,thiazole, thiophene, pyrole, pyrrolidine, pyrazole, and imidazole; or R¹is

and R² is

or is selected from the group consisting of a substituted orunsubstituted cyclic or heterocycle selected from the group consistingof pyridine, pyrimidine, naphthalene, quinolone, isoquinoline,cinnoline, phenyl, substituted phenyl, oxazole, furan, pyran, isoxazole,thiazole, thiophene, pyrole, pyrrolidine, pyrazole, and imidazole; andR³ is selected from the group consisting of H, CH₃, ethyl, propyl,butyl, CF₃, NH₂, halogen, and CH₂O where R is H, alkyl (e.g., C1-C6carbon chain), or aryl (e.g., phenyl, substituted phenyl, orheteroaryl).

Embodiment 2

The compound of embodiment 1, wherein said compound is not one or morecompounds selected from the group consisting of J03, J04, J05, J08, andJ17.

Embodiment 3

The compound of embodiment 2, wherein said compound is not J03.

Embodiment 4

The compound according to any one of embodiments 2-3, wherein saidcompound is not J04.

Embodiment 5

The compound according to any one of embodiments 2-4, wherein saidcompound is not J05.

Embodiment 6

The compound according to any one of embodiments 2-5, wherein saidcompound is not J06.

Embodiment 7

The compound according to any one of embodiments 2-6, wherein saidcompound is not J17.

Embodiment 8

The compound according to any one of embodiments 1-7, wherein R¹ is

Embodiment 9

The compound of embodiment 8, wherein R² is

where R⁴, R⁵, and R⁶ are independently selected from the groupconsisting of H, OH, halogen, methyl, and OCH₃, CF₃, ethyl, aryl, SR,SO₂R, NHCOR, and CO₂R, where R is H, alkyl (e.g., C1-C6 carbon chain),or aryl (e.g., phenyl, substituted phenyl, or heteroaryl).

Embodiment 10

The compound of embodiment 1, wherein R² is

Embodiment 11

The compound of embodiment 10, wherein R¹ is

where R⁴, R⁵, and R⁶ are independently selected from the groupconsisting of H, OH, halogen, methyl, OCH₃, OCF₃, OCHF₂, N(CH₃)₂, ethyl,propyl, butyl, NH-alkyl, O-alkyl, and SO₂CH₃.

Embodiment 12

The compound of embodiment 11, where R⁴, R⁵, and R⁶ are independentlyselected from the group consisting of H, OH, halogen, methyl, and OCH₃.

Embodiment 13

The compound according to any one of embodiments 8-12, wherein R⁵ isOCH₃.

Embodiment 14

The compound of embodiment 13, wherein R⁴ is CH₃.

Embodiment 15

The compound of embodiment 13, wherein R⁴ is CH₃ and R⁶ is H.

Embodiment 16

The compound of embodiment 13, wherein R⁴ is OCH₃.

Embodiment 17

The compound of embodiment 13, wherein R⁴ is OCH₃ and R⁶ is H.

Embodiment 18

The compound according to any one of embodiments 8-12, wherein R⁵ isCH₃.

Embodiment 19

The compound of embodiment 18, wherein R⁴ is CH₃.

Embodiment 20

The compound of embodiment 18, wherein R⁴ is CH₃ and R⁶ is H.

Embodiment 21

The compound of embodiment 18, wherein R⁴ is CH₃ and

R⁶ is CH₃.

Embodiment 22

The compound according to any one of embodiments 8-12, wherein R⁵ ishalogen.

Embodiment 23

The compound of embodiment 22, wherein R⁵ is F or Cl.

Embodiment 24

The compound according to any one of embodiments 22-23, wherein R⁴ isCH₃.

Embodiment 25

The compound according to any one of embodiments 22-23, wherein R⁴ isCH₃ and R⁶ is CH₃.

Embodiment 26

The compound according to any one of embodiments 22-23, wherein R⁴ ishalogen.

Embodiment 27

The compound of embodiment 26, wherein R⁴ is F.

Embodiment 28

The compound of embodiment 26, wherein R⁴ is Cl.

Embodiment 29

The compound according to any one of embodiments 22-28, wherein R⁶ is H.

Embodiment 30

The compound according to any one of embodiments 22-28, wherein R⁶ isCH₃.

Embodiment 31

The compound of embodiment 8, wherein R² is

where R⁷ and R⁸ are independently H, CH₃, OCH₃, and halogen.

Embodiment 32

The compound of embodiment 10, wherein R¹ is

where R⁷ and R⁸ are independently H, CH₃, OCH₃, and halogen.

Embodiment 33

The compound according to any one of embodiments 31-32, wherein R⁷ isCH₃ and R⁸ is CH₃.

Embodiment 34

The compound of embodiment 8, wherein R² is

Embodiment 35

The compound of embodiment 10, wherein R¹ is

Embodiment 36

The compound of embodiment 8, wherein R² is

Embodiment 37

The compound of embodiment 10, wherein R¹ is

Embodiment 38

The compounds according to any one of embodiments 1-37, wherein R⁰ isNH.

Embodiment 39

The compound according to any one of embodiments 31-33, wherein R⁰ isabsent.

Embodiment 40

The compound of embodiment 1, wherein said compound is a compoundaccording to a formula selected from the group consisting of

or a pharmaceutically acceptable salt or solvate thereof.

Embodiment 41

The compound of embodiment 1, wherein said compound is a compoundaccording to a formula selected from the group consisting of J14, J19,J20, J21, J22, J23, J24, and J25, or a pharmaceutically acceptable saltor solvate thereof.

Embodiment 42

The composition according to any one of embodiments 1-41 wherein saidcomposition is effective to decrease in corticotropin-releasing factor(CRF-1) induced p-tau.

Embodiment 43

The compound according to any one of embodiments 1-42, wherein saidcompound is a substantially pure S enantiomer.

Embodiment 44

The compound according to any one of embodiments 1-42, wherein saidcompound is a substantially pure R enantiomer.

Embodiment 45

A pharmaceutical formulation comprising a compound according to any oneof embodiments 1-44 and a pharmaceutically acceptable carrier orexcipient.

Embodiment 46

The formulation of embodiment 45, wherein said formulation is a unitdosage formulation.

Embodiment 47

The formulation according to any one of embodiments 45-46, wherein saidcomposition is formulated for administration via a route selected fromthe group consisting of isophoretic delivery, transdermal delivery,aerosol administration, administration via inhalation, oraladministration, intravenous administration, and rectal administration.

Embodiment 48

A method of decreasing p-tau in a mammal or inhibiting or preventing anincrease in p-tau in a mammal, said method comprising:

administering to said mammal an effective amount of one or morecompounds according to any one of embodiments 1-44; and/or

administering to said mammal an effective amount of a compound selectedfrom the group consisting of J03, J04, J05, J08, and J17, or apharmaceutically acceptable salt or solvate thereof; and/or

administering to said mammal an effective amount of a formulationaccording to any one of embodiments 45-47; and/or

administering to said mammal an effective amount of a formulationcomprising a compound selected from the group consisting of J03, J04,J05, J08, and J17 and a pharmaceutically acceptable carrier orexcipient.

Embodiment 49

A method of promoting the processing of amyloid precursor protein (APP)by the non-amyloidogenic pathway in a mammal, said method comprising:

administering to said mammal an effective amount of one or morecompounds according to any one of embodiments 1-44; and/or

administering to said mammal an effective amount of a compound selectedfrom the group consisting of J03, J04, J05, J08, and J17, or apharmaceutically acceptable salt or solvate thereof; and/or

administering to said mammal an effective amount of a formulationaccording to any one of embodiments 45-47; and/or

administering to said mammal an effective amount of a formulationcomprising a compound selected from the group consisting of J03, J04,J05, J08, and J17 and a pharmaceutically acceptable carrier orexcipient.

Embodiment 50

The method of embodiment 49, wherein said method decreases p-tau in saidmammal.

Embodiment 51

A method of preventing or delaying the onset of a pre-Alzheimer'scondition and/or cognitive dysfunction, and/or ameliorating one or moresymptoms of a pre-Alzheimer's condition and/or cognitive dysfunction, orpreventing or delaying the progression of a pre-Alzheimer's condition orcognitive dysfunction to Alzheimer's disease in a mammal, said methodcomprising:

administering to said mammal an effective amount of one or morecompounds according to any one of embodiments 1-44; and/or

administering to said mammal an effective amount of a compound selectedfrom the group consisting of J03, J04, J05, J08, and J17, or apharmaceutically acceptable salt or solvate thereof; and/or

administering to said mammal an effective amount of a formulationaccording to any one of embodiments 45-47; and/or

administering to said mammal an effective amount of a formulationcomprising a compound selected from the group consisting of J03, J04,J05, J08, and J17 and a pharmaceutically acceptable carrier orexcipient.

Embodiment 52

A method of ameliorating one or more symptoms of Alzheimer's disease,and/or reversing Alzheimer's disease, and/or reducing the rate ofprogression of Alzheimer's disease in a mammal, said method comprising:

administering to said mammal an effective amount of one or morecompounds according to any one of embodiments 1-44; and/or

administering to said mammal an effective amount of a compound selectedfrom the group consisting of J03, J04, J05, J08, and J17, or apharmaceutically acceptable salt or solvate thereof; and/or

administering to said mammal an effective amount of a formulationaccording to any one of embodiments 45-47; and/or

administering to said mammal an effective amount of a formulationcomprising a compound selected from the group consisting of J03, J04,J05, J08, and J17 and a pharmaceutically acceptable carrier orexcipient.

Embodiment 53

The method according to any one of embodiments 48-52, wherein saidmethod comprises administering a compound selected from the groupconsisting of J03, J04, J05, J06, J07, J08, J09, J10, J11, J12, J14,J15, J17, J19, J20, J21, J22, J23, J24, and J25, or a pharmaceuticallyacceptable salt or solvate thereof.

Embodiment 54

The method according to any one of embodiments 48-52, wherein saidmethod comprises administering a compound selected from the groupconsisting of J14, J19, J20, J21, J22, J23, J24, and J25, or apharmaceutically acceptable salt or solvate thereof.

Embodiment 55

The method according to any one of embodiments 48-52, wherein saidmethod comprises administering a compound selected from the groupconsisting of J03, J04, J05, J08, and J17, or a pharmaceuticallyacceptable salt or solvate thereof.

Embodiment 56

The method according to any one of embodiments 48-52, wherein saidmethod comprises administering J03, or a pharmaceutically acceptablesalt or solvate thereof.

Embodiment 57

The method according to any one of embodiments 48-56, wherein the mammalhas a familial risk for having Alzheimer's disease.

Embodiment 58

The method according to any one of embodiments 48-56, wherein the mammalhas a familial Alzheimer's disease (FAD) mutation.

Embodiment 59

The method according to any one of embodiments 48-56, wherein saidmammal has one copy of the ApoE4 allele.

Embodiment 60

The method according to any one of embodiments 48-56, wherein saidmammal has two copies of the ApoE4 allele.

Embodiment 61

The method according to any one of embodiments 48-60, wherein saidmammal is a human.

Embodiment 62

The method according to any one of embodiments 48-61, wherein, whereinsaid method is a method of preventing or delaying the transition from acognitively asymptomatic pre-Alzheimer's condition to a pre-Alzheimer'scognitive dysfunction.

Embodiment 63

The method according to any one of embodiments 48-61, wherein saidmethod is a method of preventing or delaying the onset of apre-Alzheimer's cognitive dysfunction.

Embodiment 64

The method according to any one of embodiments 48-63, wherein saidmethod comprises ameliorating one or more symptoms of a pre-Alzheimer'scognitive dysfunction.

Embodiment 65

The method according to any one of embodiments 48-63, wherein saidmethod comprises preventing or delaying the progression of apre-Alzheimer's cognitive dysfunction to Alzheimer's disease.

Embodiment 66

The method of embodiment 65, wherein said method delays or prevents theprogression of MCI to Alzheimer's disease.

Embodiment 67

The method according to any one of embodiments 48-65, wherein saidmammal exhibits biomarker positivity of Aβ in a clinically normal humanmammal age 50 or older.

Embodiment 68

The method according to any one of embodiments 48-65, wherein saidmammal exhibits asymptomatic cerebral amyloidosis.

Embodiment 69

The method according to any one of embodiments 48-65, wherein saidmammal exhibits cerebral amyloidosis in combination with downstreamneurodegeneration.

Embodiment 70

The method according to any one of embodiments 48-65, wherein saidmammal is cognitively asymptomatic.

Embodiment 71

The method according to any one of embodiments 48-65, wherein saidmammal exhibits cerebral amyloidosis in combination with downstreamneurodegeneration and subtle cognitive/behavioral decline.

Embodiment 72

The method of embodiment 71, wherein said downstream neurodegenerationis determined by one or more elevated markers of neuronal injuryselected from the group consisting of tau, and FDG uptake.

Embodiment 73

The method according to any one of embodiments 68-72, wherein saidcerebral amyloidosis is determined by PET, or CSF analysis, andstructural MRI (sMRI).

Embodiment 74

The method according to any one of embodiments 48-65, wherein saidmammal is a mammal diagnosed with mild cognitive impairment.

Embodiment 75

The method of embodiment 74, wherein said mammal shows a clinicaldementia rating above zero and below about 1.5.

Embodiment 76

The method according to any one of embodiments 48-66, wherein the mammalis not diagnosed as at risk for a neurological disease or disorder otherthan Alzheimer's disease.

Embodiment 77

The method according to any one of embodiments 48-76, wherein saidadministration produces a reduction in the CSF of levels of one or morecomponents selected from the group consisting of total-Tau (tTau),phospho-Tau (pTau), APPneo, soluble Aβ40, pTau/Aβ42 ratio and tTau/Aβ42ratio, and/or an increase in the CSF of levels of one or more componentsselected from the group consisting of Aβ42/Aβ40 ratio, Aβ42/Aβ38 ratio,sAPPα, sAPPα/sAPPβ ratio, sAPPα/Aβ40 ratio, and sAPPα/Aβ42 ratio.

Embodiment 78

The method according to any one of embodiments 48-77, wherein saidadministration produces a reduction of the plaque load in the brain ofthe mammal.

Embodiment 79

The method according to any one of embodiments 48-78, wherein saidadministration produces a reduction in the rate of plaque formation inthe brain of the mammal.

Embodiment 80

The method according to any one of embodiments 48-79, wherein saidadministration produces an improvement in the cognitive abilities of themammal.

Embodiment 81

The method according to any one of embodiments 48-79, wherein saidadministration produces an improvement in, a stabilization of, or areduction in the rate of decline of the clinical dementia rating (CDR)of the mammal.

Embodiment 82

The method according to any one of embodiments 48-81, wherein the mammalis a human and said administration produces a perceived improvement inquality of life by the human.

Embodiment 83

The method according to any one of embodiments 48-82, wherein thecompound(s) are administered via a route selected from the groupconsisting of oral delivery, isophoretic delivery, transdermal delivery,parenteral delivery, aerosol administration, administration viainhalation, intravenous administration, and rectal administration.

Embodiment 84

The method according to any one of embodiments 48-82, wherein thecompound is administered orally.

Embodiment 85

The method according to any one of embodiments 48-84, wherein theadministering is over a period of at least three weeks.

Embodiment 86

The method according to any one of embodiments 48-84, wherein theadministering is over a period of at least 6 months.

Embodiment 87

The method according to any one of embodiments 48-86, wherein thecompound(s) are administered via a route selected from the groupconsisting of isophoretic delivery, transdermal delivery, aerosoladministration, administration via inhalation, oral administration,intravenous administration, and rectal administration.

Definitions

Unless otherwise indicated, reference to a compound (e.g., to atriazolopyrimidine and/or triazolopyridine as described herein) shouldbe construed broadly to include pharmaceutically acceptable salts,prodrugs, tautomers, alternate solid forms, non-covalent complexes, andcombinations thereof, of a chemical entity of the depicted structure orchemical name.

Generally, reference to a certain element such as hydrogen or H is meantto include all isotopes of that element. For example, if an R group isdefined to include hydrogen or H, it also includes deuterium andtritium. Accordingly, isotopically labeled compounds are within thescope of this invention.

A pharmaceutically acceptable salt is any salt of the parent compoundthat is suitable for administration to an animal or human. Apharmaceutically acceptable salt also refers to any salt which may formin vivo as a result of administration of an acid, another salt, or aprodrug which is converted into an acid or salt. A salt comprises one ormore ionic forms of the compound, such as a conjugate acid or base,associated with one or more corresponding counterions. Salts can formfrom or incorporate one or more deprotonated acidic groups (e.g.carboxylic acids), one or more protonated basic groups (e.g. amines), orboth (e.g. zwitterions).

A prodrug is a compound that is converted to a therapeutically activecompound after administration. For example, conversion may occur byhydrolysis of an ester group, such as a C₁-C₆ alkyl ester of thecarboxylic acid group of the present compounds, or some otherbiologically labile group. Prodrug preparation is well known in the art.For example, “Prodrugs and Drug Delivery Systems,” which is a chapter inRichard B. Silverman, Organic Chemistry of Drug Design and Drug Action,2d Ed., Elsevier Academic Press: Amsterdam, 2004, pp. 496-557, providesfurther detail on the subject.

Tautomers are isomers that are in equilibrium with one another. Forexample, tautomers may be related by transfer of a proton, hydrogenatom, or hydride ion.

Unless stereochemistry is explicitly depicted, a structure is intendedto include every possible stereoisomer, both pure or in any possiblemixture.

Alternate solid forms are different solid forms than those that mayresult from practicing the procedures described herein. For example,alternate solid forms may be polymorphs, different kinds of amorphoussolid forms, glasses, and the like. In various embodiments alternatesolid forms of any of the compounds described herein are contemplated.

In general, “substituted” refers to an organic group as defined below(e.g., an alkyl group) in which one or more bonds to a hydrogen atomcontained therein are replaced by a bond to non-hydrogen or non-carbonatoms. Substituted groups also include groups in which one or more bondsto a carbon(s) or hydrogen(s) atom are replaced by one or more bonds,including double or triple bonds, to a heteroatom. Thus, a substitutedgroup will be substituted with one or more substituents, unlessotherwise specified. In some embodiments, a substituted group issubstituted with 1, 2, 3, 4, 5, or 6 substituents. Examples ofsubstituent groups include: halogens (i.e., F, Cl, Br, and I);hydroxyls; alkoxy, alkenoxy, alkynoxy, aryloxy, aralkyloxy,heterocyclyloxy, and heterocyclylalkoxy groups; carbonyls (oxo);carboxyls; esters; urethanes; oximes; hydroxylamines; alkoxyamines;aralkoxyamines; thiols; sulfides; sulfoxides; sulfones; sulfonyls;sulfonamides; amines; N-oxides; hydrazines; hydrazides; hydrazones;azides; amides; ureas; amidines; guanidines; enamines; imides;isocyanates; isothiocyanates; cyanates; thiocyanates; imines; nitrogroups; nitriles (i.e., CN), and the like.

The term “alkyl” refers to and covers any and all groups that are knownas normal alkyl, branched-chain alkyl, cycloalkyl and alsocycloalkyl-alkyl. Illustrative alkyl groups include, but are not limitedto methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl,t-butyl, octyl, and decyl. The term “cycloalkyl” refers to cyclic,including polycyclic, saturated hydrocarbyl groups. Examples include,but are not limited to cyclopentyl, cyclohexyl, dicyclopentyl,norbornyl, octahydronapthyl, and spiro[3.4]octyl. In certainembodiments, alkyl groups contain 1-12 carbon atoms (C1-12 alkyl), or1-9 carbon atoms (C₁₋₉ alkyl), or 1-6 carbon atoms (C₁₋₆ alkyl), or 1-5carbon atoms (C₁₋₅ alkyl), or carbon atoms (C₁₋₄ alkyl), or 1-3 carbonatoms (C₁₋₃ alkyl), or 1-2 carbon atoms (C₁₋₂ alkyl).

By way of example, the term “C₁₋₆ alkyl group” refers to a straightchain or branched chain alkyl group having 1 to 6 carbon atoms, and maybe exemplified by a methyl group, an ethyl group, an n-propyl group, anisopropyl group, an n-butyl group, an isobutyl group, a tert-butylgroup, a sec-butyl group, an n-pentyl group, a tert-amyl group, a3-methylbutyl group, a neopentyl group, and an n-hexyl group.

The term “alkoxy” as used herein means an alkyl group bound through asingle, terminal oxygen atom. An “alkoxy” group may be represented as—O-alkyl where alkyl is as defined above. The term “aryloxy” is used ina similar fashion, and may be represented as —O-aryl, with aryl asdefined below. The term “hydroxy” refers to —OH.

Similarly, the term “alkylthio” as used herein means an alkyl groupbound through a single, terminal sulfur atom. An “alkylthio” group maybe represented as —S-alkyl where alkyl is as defined above. The term“arylthio” is used similarly, and may be represented as —S-aryl, witharyl as defined below. The term “mercapto” refers to —SH.

Aryl groups are cyclic aromatic hydrocarbons that do not containheteroatoms. Aryl groups include monocyclic, bicyclic and polycyclicring systems. Thus, aryl groups include, but are not limited to, phenyl,azulenyl, heptalenyl, biphenylenyl, indacenyl, fluorenyl, phenanthrenyl,triphenylenyl, pyrenyl, naphthacenyl, chrysenyl, biphenyl, anthracenyl,indenyl, indanyl, pentalenyl, and naphthyl groups. In some embodiments,aryl groups contain 6-14 carbons, and in others from 6 to 12 or even6-10 carbon atoms in the ring portions of the groups. Although thephrase “aryl groups” includes groups containing fused rings, such asfused aromatic-aliphatic ring systems (e.g., indanyl,tetrahydronaphthyl, and the like), it does not include aryl groups thathave other groups, such as alkyl or halo groups, bonded to one of thering members. Rather, groups such as tolyl are referred to assubstituted aryl groups. Representative substituted aryl groups may bemono-substituted or substituted more than once. For example,monosubstituted aryl groups include, but are not limited to, 2-, 3-, 4-,5-, or 6-substituted phenyl or naphthyl groups, which may be substitutedwith substituents such as those listed above.

The term “heteroaryl group” refers to a monocyclic or condensed-ringaromatic heterocyclic group containing one or more hetero-atoms selectedfrom O, S and N. If the aromatic heterocyclic group has a condensedring, it can include a partially hydrogenated monocyclic group. Examplesof such a heteroaryl group include a pyrazolyl group, a thiazolyl group,an isothiazolyl group, a thiadiazolyl group, an imidazolyl group, afuryl group, a thienyl group, an oxazolyl group, an isoxazolyl group, apyrrolyl group, an imidazolyl group, a (1,2,3)- and (1,2,4)-triazolylgroup, a tetrazolyl group, a pyranyl group, a pyridyl group, apyrimidinyl group, a pyrazinyl group, a pyridazinyl group, a quinolylgroup, an isoquinolyl group, a benzofuranyl group, an isobenzofuranylgroup, an indolyl group, an isoindolyl group, an indazolyl group, abenzoimidazolyl group, a benzotriazolyl group, a benzoxazolyl group, abenzothiazolyl group, a benzo[b]thiophenyl group, athieno[2,3-b]thiophenyl group, a (1,2)- and (1,3)-benzoxathiol group, achromenyl group, a 2-oxochromenyl group, a benzothiadiazolyl group, aquinolizinyl group, a phthalazinyl group, a naphthyridinyl group, aquinoxalinyl group, a quinazolinyl group, a cinnolinyl group, and acarbazolyl group.

A “derivative” of a compound means a chemically modified compoundwherein the chemical modification takes place at one or more functionalgroups of the compound. The derivative however, is expected to retain,or enhance, the pharmacological activity of the compound from which itis derived.

As used herein, “administering” refers to local and systemicadministration, e.g., including enteral, parenteral, pulmonary, andtopical/transdermal administration. Routes of administration for agents(e.g., triazolopyrimidines and/or triazolopyridines described herein, ora tautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts or solvates of said compound(s), said stereoisomer(s), or saidtautomer(s), or analogues, derivatives, or prodrugs thereof) that finduse in the methods described herein include, e.g., oral (per os (p.o.))administration, nasal or inhalation administration, administration as asuppository, topical contact, transdermal delivery (e.g., via atransdermal patch), intrathecal (IT) administration, intravenous (“iv”)administration, intraperitoneal (“ip”) administration, intramuscular(“im”) administration, intralesional administration, or subcutaneous(“sc”) administration, or the implantation of a slow-release devicee.g., a mini-osmotic pump, a depot formulation, etc., to a subject.Administration can be by any route including parenteral and transmucosal(e.g., oral, nasal, vaginal, rectal, or transdermal). Parenteraladministration includes, e.g., intravenous, intramuscular,intra-arterial, intradermal, subcutaneous, intraperitoneal,intraventricular, ionophoretic and intracranial. Other modes of deliveryinclude, but are not limited to, the use of liposomal formulations,intravenous infusion, transdermal patches, etc.

The terms “systemic administration” and “systemically administered”refer to a method of administering the agent(s) described herein orcomposition to a mammal so that the agent(s) or composition is deliveredto sites in the body, including the targeted site of pharmaceuticalaction, via the circulatory system. Systemic administration includes,but is not limited to, oral, intranasal, rectal and parenteral (e.g.,other than through the alimentary tract, such as intramuscular,intravenous, intra-arterial, transdermal and sub cutaneous)administration.

The term “co-administering” or “concurrent administration” or“administering in conjunction with” when used, for example with respectto the active agent(s) described herein e.g., triazolopyrimidine(s)and/or triazolopyridine(s) described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts orsolvates of said compound(s), said stereoisomer(s), or said tautomer(s),or analogues, derivatives, or prodrugs thereof and a second active agent(e.g., a cognition enhancer), refers to administration of the agent(s)and/the second active agent such that both can simultaneously achieve aphysiological effect. The two agents, however, need not be administeredtogether. In certain embodiments, administration of one agent canprecede administration of the other. Simultaneous physiological effectneed not necessarily require presence of both agents in the circulationat the same time. However, in certain embodiments, co-administeringtypically results in both agents being simultaneously present in thebody (e.g., in the plasma) at a significant fraction (e.g., 20% orgreater, preferably 30% or 40% or greater, more preferably 50% or 60% orgreater, most preferably 70% or 80% or 90% or greater) of their maximumserum concentration for any given dose.

The term “effective amount” or “pharmaceutically effective amount” referto the amount and/or dosage, and/or dosage regime of one or moreagent(s) necessary to bring about the desired result e.g., an amountsufficient to mitigating in a mammal one or more symptoms associatedwith mild cognitive impairment (MCI), or an amount sufficient to lessenthe severity or delay the progression of a disease characterized byamyloid deposits in the brain in a mammal (e.g., therapeuticallyeffective amounts), an amount sufficient to reduce the risk or delayingthe onset, and/or reduce the ultimate severity of a diseasecharacterized by amyloid deposits in the brain in a mammal (e.g.,prophylactically effective amounts).

The phrase “cause to be administered” refers to the actions taken by amedical professional (e.g., a physician), or a person controllingmedical care of a subject, that control and/or permit the administrationof the agent(s) at issue to the subject. Causing to be administered caninvolve diagnosis and/or determination of an appropriate therapeutic orprophylactic regimen, and/or prescribing particular agent(s) for asubject. Such prescribing can include, for example, drafting aprescription form, annotating a medical record, and the like.

As used herein, the terms “treating” and “treatment” refer to delayingthe onset of, retarding or reversing the progress of, reducing theseverity of, or alleviating or preventing either the disease orcondition to which the term applies, or one or more symptoms of suchdisease or condition.

The term “mitigating” refers to reduction or elimination of one or moresymptoms of that pathology or disease, and/or a reduction in the rate ordelay of onset or severity of one or more symptoms of that pathology ordisease, and/or the prevention of that pathology or disease. In certainembodiments, the reduction or elimination of one or more symptoms ofpathology or disease can include, but is not limited to, reduction orelimination, or prevention of an increase (e.g., a stress-inducedincrease), of one or more markers that are characteristic of thepathology or disease (e.g., of total-Tau (tTau), phospho-Tau (pTau),APPneo, soluble Aβ40, pTau/Aβ42 ratio and tTau/Aβ42 ratio, and/or anincrease in the CSF of levels of one or more components selected fromthe group consisting of Aβ42/Aβ40 ratio, Aβ42/Aβ38 ratio, sAPPα,sAPPα/sAPPβ ratio, sAPPα/Aβ40 ratio, sAPPα/Aβ42 ratio, etc.) and/orreduction, stabilization or reversal of one or more diagnostic criteria(e.g., clinical dementia rating (CDR)).

As used herein, the phrase “consisting essentially of” refers to thegenera or species of active pharmaceutical agents recited in a method orcomposition, and further can include other agents that, on their own donot substantial activity for the recited indication or purpose. In someembodiments, the phrase “consisting essentially of” expressly excludesthe inclusion of one or more additional agents that haveneuropharmacological activity other than the recited agent(s) (e.g.,other than ASBIs such as galangin, rutin, and analogues, derivatives, orprodrugs thereof). In some embodiments, the phrase “consistingessentially of” expressly excludes the inclusion of one or moreadditional active agents other than the active agent(s) described herein(e.g., other than ASBIs such as galangin, rutin, and analogues,derivatives, or prodrugs thereof). In some embodiments, the phrase“consisting essentially of” expressly excludes the inclusion of one ormore acetylcholinesterase inhibitors.

The terms “subject”, “individual”, and “patient” interchangeably referto a mammal, preferably a human or a non-human primate, but alsodomesticated mammals (e.g., canine or feline), laboratory mammals (e.g.,mouse, rat, rabbit, hamster, guinea pig) and agricultural mammals (e.g.,equine, bovine, porcine, ovine). In various embodiments, the subject canbe a human (e.g., adult male, adult female, adolescent male, adolescentfemale, male child, female child) under the care of a physician or otherhealth worker in a hospital, psychiatric care facility, as anoutpatient, or other clinical context. In certain embodiments thesubject may not be under the care or prescription of a physician orother health worker.

The term “formulation” or “drug formulation” or “dosage form” or“pharmaceutical formulation” as used herein refers to a compositioncontaining at least one therapeutic agent or medication for delivery toa subject. In certain embodiments the dosage form comprises a given“formulation” or “drug formulation” and may be administered to a patientin the form of a lozenge, pill, tablet, capsule, suppository, membrane,strip, liquid, patch, film, gel, spray or other form.

The term “mucosal membrane” refers generally to any of the mucus-coatedbiological membranes in the body. In certain embodiments active agent(s)described herein can be administered herein via any mucous membranefound in the body, including, but not limited to buccal, perlingual,nasal, sublingual, pulmonary, rectal, and vaginal mucosa. Absorptionthrough the mucosal membranes of the oral cavity and those of the gutare of interest. Thus, peroral, buccal, sublingual, gingival and palatalabsorption are contemplated herein.

The term “transmucosal” delivery of a drug and the like is meant toencompass all forms of delivery across or through a mucosal membrane.

The term “bioadhesion” as used herein refers to the process of adhesionof the dosage form(s) to a biological surface, e.g., mucosal membranes.

“Controlled drug delivery” refers to release or administration of a drugfrom a given dosage form in a controlled fashion in order to achieve thedesired pharmacokinetic profile in vivo. An aspect of “controlled” drugdelivery is the ability to manipulate the formulation and/or dosage formin order to establish the desired kinetics of drug release.

“Sustained drug delivery” refers to release or administration of a drugfrom a source (e.g., a drug formulation) in a sustained fashion over aprotracted yet specific period of time, that may extend from severalminutes to a few hours, days, weeks or months. In various embodimentsthe term “sustained” will be used to refer to delivery of consistentand/oe substantially constant levels of drug over a time period rangingfrom a few minutes to a day, with a profile characterized by the absenceof an immediate release phase, such as the one obtained from IVadministration.

The term “T_(max)” as used herein means the time point of maximumobserved plasma concentration.

The term “C_(max)” as used herein means the maximum observed plasmaconcentration.

The term “plasma t_(1/2)” as used herein means the observed “plasmahalf-life” and represents the time required for the drug plasmaconcentration to reach the 50% of its maximal value (C_(max)). Thisfacilitates determination of the mean duration of pharmacologicaleffects. In addition, it facilitates direct and meaningful comparisonsof the duration of different test articles after delivery via the sameor different routes.

The term “Optimal Therapeutic Targeting Ratio” or “OTTR” represents theaverage time that the drug is present at therapeutic levels, defined astime within which the drug plasma concentration is maintained above 50%of C_(max) normalized by the drug's elimination half-life multiplied bythe ratio of the C_(max) obtained in the dosage form of interest overthe C_(max) following IV administration of equivalent doses and it iscalculated by the formula:

OTTR=(C ^(IV) _(max) /C _(max))×(Dose/Dose^(IV))(Time above 50% of C_(max))/(Terminal^(IV) elimination half-life of the drug).

The term “substantially pure” means sufficiently homogeneous to appearfree of readily detectable impurities as determined by standard methodsof analysis, such as thin layer chromatography (TLC), gelelectrophoresis and high performance liquid chromatography (HPLC), usedby those of skill in the art to assess such purity, or sufficiently puresuch that further purification would not detectably alter the physicalor chemical properties, of the compound. Methods for purification of thecompounds to produce substantially chemically pure compounds are knownto those of skill in the art. A substantially chemically pure compoundmay, however, be a mixture of stereoisomers or isomers. In suchinstances, further purification might increase the specific activity ofthe compound.

The term “substantially pure” when used with respect to enantiomersindicates that one particular enantiomer (e.g. an S enantiomer or an Renantiomer) is substantially free of its stereoisomer. In variousembodiments substantially pure indicates that a particular enantiomer isat least 70%, or at least 80%, or at least 90%, or at least 95%, or atleast 98%, or at least 99% of the purified compound. Methods ofproducing substantially pure enantiomers are well known to those ofskill in the art. For example, a single stereoisomer, e.g., anenantiomer, substantially free of its stereoisomer may be obtained byresolution of the racemic mixture using a method such as formation ofdiastereomers using optically active resolving agents (Stereochemistryof Carbon Compounds, (1962) by E. L. Eliel, McGraw Hill; Lochmuller(1975) J Chromatogr., 113(3): 283-302). Racemic mixtures of chiralcompounds of the can be separated and isolated by any suitable method,including, but not limited to: (1) formation of ionic, diastereomericsalts with chiral compounds and separation by fractional crystallizationor other methods, (2) formation of diastereomeric compounds with chiralderivatizing reagents, separation of the diastereomers, and conversionto the pure stereoisomers, and (3) separation of the substantially pureor enriched stereoisomers directly under chiral conditions. Anotherapproach for separation of the enantiomers is to use a Diacel chiralcolumn and elution using an organic mobile phase such as done by ChiralTechnologies (www.chiraltech.com) on a fee for service basis.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates design considerations in making J03 analogs.

FIG. 2 illustrates a number of triazolopyridines and triazolopyrimidinesthat lower stress-induced p-tau.

FIG. 3 illustrates a synthesis Scheme 1 for J19. Reagents andconditions: (a) 2-methoxyethanol, 125° C.; (b) NaNO₂, DCM, acetic acid,r.t; (c) THF, 100° C., sealed tube.

FIG. 4, panels A-C, show that J03 (panel A) reduces tau (panel B) andphospho-tau (panel C) increases induced by CRF. SHSY-5Y cells werecultured without serum to induce differentiation and increased tauexpression.

FIG. 5 Illustrates in vivo pharmacokinetics of J03 (10 mk delivered in asingle 5 mg/ml DMSO stock, 60 subcutaneous injection).

FIG. 6, panels A-F, illustrate non-tau data resulting from initial J03study. Panel A: Familiarity; Panel B: Aβ1-40; Panel C: Aβ1-42; Panel D:Aβ1-40/Aβ1-42; Panel E: sAPPα levels; Panel F: sAPPα/Aβ1-42.

FIG. 7, illustrates effect of J03 on tau (top left), p-tau (top right),and the ratio p-tau/tau (bottom left).

FIG. 8 illustrates the performance of J03-treated mice in both novellocation (left) and object (right) assays.

FIG. 9 illustrates the effect of J03 on sAPPα (top left), sAPPβ (topright), and the sAPPα/sAPPβ ratio (bottom left).

FIG. 10 illustrates the effect of J03 on Aβ1-40 (top left), Aβ1-42 (topright), and the Aβ1-40/Aβ1-42 ratio (bottom left).

FIG. 11 illustrates the effect of J03 on Aβ1-42 in two studies.

FIG. 12 illustrates the effect of J03 on phosphorylated tau (p-tau).

FIG. 13, panels A-F, illustrates the effect of J03 on p-tau and memory.Panel A: Novel object preference; Panel B: p-tau versus novel objectpreference; Panel C: Relationship between novel object preference (NOP)score and p-tau; Panel D: Effect of J03 on novel location preference(NLP); Panel E p-tau versus novel location preference; Panel F:Relationship between novel location preference (NLP) score and p-tau.

FIG. 14 illustrates plasma and brain levels of J03 two hours after oraldosing (left panel) and at the end of the study (right panel).

FIG. 15 illustrates the effect of J03, J04, J07, and J12 on tau andp-tau.

FIG. 16 illustrates the pharmacokinetics of J04 in brain and plasmaafter injection and oral delivery (left panel) or after oral delivery(right panel).

FIG. 17 schematically illustrates the reversal of substituents aroundthe triazolopyrimidine ring. With respect to any of the compoundsdescribed herein the reversed substituent form is contemplated as well.

FIG. 18 illustrates the effect of J17 on tau (left panel) and p-tau(right panel) in SH-SY5Y cells.

FIG. 19 illustrates the effect of J17 on sAPPα (top left), tau (topright), and p-tau (bottom left) after CRF stimulation.

FIG. 20 illustrates the effect J03 and J17 on sAppα (top left), tau (topright), and p-tau (bottom left) with increasing concentrations of CRF.

FIG. 21 illustrates in vivo pharmacokinetics of J17.

FIG. 22 illustrates the effect of J03 and J17 on total interactions andnovel object preference.

FIG. 23 illustrates the effect of J03 and J17 on sAPPα (top panel),sAPPβ (middle panel) and the ratio (bottom panel) broken out by gender.

FIG. 24 illustrates the effect of J03 and J17 on Aβ1-42.

FIG. 25 illustrates the effect of J03 and J17 on the Aβ1-40/Aβ1-42 ratiobroken out by gender with outliers removed.

FIG. 26 illustrates the effect of J03 and J17 on the sAPPα/Aβ1-42 ratiobroken out by gender with outliers included (top panel) and withoutliers removed (bottom panel).

FIG. 27 illustrates the effects of J03 and J17 on the values of tau (toppanel), p-tau (middle panel), and the ration p-tau/tau (lower panel)broken out by gender.

FIG. 28, panels A-D illustrate the effect of J03 and J19 on CRF-inducedtau and p-tau alterations. Panel A: tau; Panel B: p-tau; Panel C:p-tau/tau ratio; Panel D: sAppα.

FIG. 29 illustrates the effect of J19 on tau (left panel), p-tau (leftpanel), and the ratio (right panel).

FIG. 30 illustrates the in vivo pharmacokinetics of J19.

DETAILED DESCRIPTION

Novel compounds that inhibit corticotropin-releasing factor CRF-1associated phosphorylation of tau are identified herein. Without beingbound to a particular theory, it is believed these compounds showefficacy in the treatment of ongoing Alzheimer's Disease, in the delayor prevention of the onset of Alzheimer's disease, in the onset of mildcognitive impairment (MCI) when mediated by an amyloidogenic process, inthe delay of a transition from MCI to AD, and in the delay or preventionof MCI.

In people diagnosed with Alzheimer's disease (AD), and in our handsusing the J20 mouse model of AD, the level of tau phosphorylationprovides the closest correlation to degree of cognitive impairment. Thereversal of tau pathology alone can improve memory, even in the presenceof high Aβ42 in J20 mice Stress and the associated increase incorticotropin-releasing factor CRF-1 is known to increase thephosphorylation of tau.

To identify therapeutic candidates, a clinical library of CRF-1Rinhibitors to was screened to determine their effects oncortisol-induced p-tau increases. One compound, designated “J03”(N-(4-Methoxy-2-methylphenyl)-1-[1-(methoxymethyl)propyl]-6-methyl-1H-1,2,3-triazolo[4,5-c]pyridine-4-amine,see FIG. 4) acted as a CRF-1 antagonist (Ki˜7.9 nM) and showed nobinding to CRFR2 (Ki>10,000 nM). J03 was shown specifically to inhibitstress-induced p-tau increases by cortisol in vitro. Notably, testing ofanother set of CRF1 antagonists did not induce a similar inhibition ofp-tau.

Following the observations of J03 a number of analogs were developed.One design focus was to replace the triazolopyridine ring of J03 with atriazolopyrimidine ring (see, e.g., FIG. 1) and to explore theorientation of and vary the substituents around the triazolopyridine andtriazolopyrimidine rings. It is noted that with respect to anytriazolopyridine and triazolopyrimidine described herein a compound withsubstituents A and B in reversed positions (see, e.g., FIG. 1) is alsocontemplated. Synthetic schemes have been developed and the analogsynthesis has been performed. Biological activity and pharmacokineticshas been evaluated (see, Example 2).

A number of compounds are illustrated in FIG. 2 and some properties ofthese compounds are summarized below in Table 1.

As illustrated, a number of these compounds are effective in loweringp-tau and/or reducing or preventing a stress-induced increase in p-tau.Moreover, as indicated above, reduction in p-tau (or inhibition of p-tauincrease) is an important metric of efficacy in pathologiescharacterized by the accumulation of amyloid plaque (e.g., Alzheimer'sdisease, MCI, etc.). It is believed these compounds and analogs thereof,pharmaceutically acceptable salts and clathrates thereof, and the likeare useful in the prophylaxis and/or treatment of pathologiescharacterized by the accumulation of amyloid plaque.

Accordingly it is believed that these agents) (e.g., triazolopyrimidineand/or triazolopyridine compounds described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts orsolvates of said compounds(s), said stereoisomer(s), or saidtautomer(s), or analogues, derivatives, or prodrugs thereof) can be usedto decrease p-tau in a mammal, and/or to inhibit or prevent an increasein p-tau, and/or to prevent or delay the onset of a pre-Alzheimer'scognitive dysfunction, and/or to ameliorate one or more symptoms of apre-Alzheimer's cognitive dysfunction, and/or to prevent or delay theprogression of a pre-Alzheimer's condition or cognitive dysfunction toAlzheimer's disease, and/or to promote the processing of amyloidprecursor protein (APP) by the non-amyloidogenic pathway. In certainembodiments these agents can be used in the treatment of Alzheimer'sdisease (e.g., to lessen the severity of the disease, and/or toameliorate one or more symptoms of the disease, and/or to slow theprogression of the disease).

While the methods described herein are detailed primarily in the contextof mild cognitive impairment (MCI) and Alzheimer's disease (AD) it isbelieved they can apply equally to other pathologies characterized byamyloidosis. In this respect, an illustrative, but non-limiting list ofconditions characterized by amyloid plaque formation is shown in Table2.

TABLE 2 Illustrative pathologies characterized by amyloidformation/deposition. Disease Characteristic Protein AbbreviationAlzheimer's disease Beta amyloid Aβ Diabetes mellitus type 2 Isletamyloid protein IAPP (Amylin) Parkinson's disease Alpha-synuclein SNCATransmissible spongiform Prion PrP encephalopathy e.g. Bovine spongiformencephalopathy Huntington's Disease Huntingtin HTT Medullary carcinomaof the thyroid Calcitonin ACal Cardiac arrhythmias, Isolated Atrialnatriuretic factor AANF atrial amyloidosis AtherosclerosisApolipoprotein AI AApoA1 Rheumatoid arthritis Serum amyloid A AA Aorticmedial amyloid Medin AMed Prolactinomas Prolactin APro Familial amyloidpolyneuropathy Transthyretin ATTR Hereditary non-neuropathic LysozymeALys systemic amyloidosis Dialysis related amyloidosis Beta 2microglobulin Aβ2M Finnish amyloidosis Gelsolin AGel Lattice cornealdystrophy Keratoepithelin AKer Cerebral amyloid angiopathy Beta amyloidAβ Cerebral amyloid angiopathy Cystatin ACys (Icelandic type) SystemicAL amyloidosis Immunoglobulin light AL chain AL Sporadic Inclusion BodyMyositis S-IBM none Age-related macular Beta amyloid Aβ degeneration(AMD) Cerebrovascular dementia Cerebrovascular amyloid CVA

Therapeutic and Prophylactic Methods.

In various embodiments therapeutic and/or prophylactic methods areprovided that utilize the active agent(s) (e.g., triazolopyrimidine(s)and/or triazolopyridines described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts orsolvates of said triazolopyrimidine(s) and/or triazolopyridine(s), saidstereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof) are provided. Typically the methods involveadministering one or more active agent(s) to a subject (e.g., to a humanin need thereof) in an amount sufficient to realize the desiredtherapeutic or prophylactic result.

Prophylaxis

In certain embodiments active agent(s) (e.g., triazolopyrimidine(s)and/or triazolopyridine(s) described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts orsolvates of said triazolopyrimidine(s) and/or triazolopyridine(s), saidstereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof) are utilized in various prophylactic contexts. Thus,for example, in certain embodiments, the active agent(s) can be used toprevent or delay the onset of a pre-Alzheimer's cognitive dysfunction,and/or to ameliorate one more symptoms of a pre-Alzheimer's conditionand/or cognitive dysfunction, and/or to prevent or delay the progressionof a pre-Alzheimer's condition and/or cognitive dysfunction toAlzheimer's disease.

Accordingly in certain embodiments, the prophylactic methods describedherein are contemplated for subjects identified as “at risk” and/or ashaving evidence of early Alzheimer's Disease (AD) pathological changes,but who do not meet clinical criteria for MCI or dementia. Without beingbound to a particular theory, it is believed that even this“preclinical” stage of the disease represents a continuum fromcompletely asymptomatic individuals with biomarker evidence suggestiveof AD-pathophysiological process(es) (abbreviated as AD-P, see, e.g.,Sperling et al. (2011) Alzheimer's & Dementia, 1-13) at risk forprogression to AD dementia to biomarker-positive individuals who arealready demonstrating very subtle decline but not yet meetingstandardized criteria for MCI (see, e.g., Albert et al. (2011)Alzheimer's and Dementia, 1-10 (doi:10.1016/j.jalz.2011.03.008).

This latter group of individuals might be classified as “not normal, notMCI” but would be can be designated “pre-symptomatic” or “pre-clinicalor “asymptomatic” or “premanifest”). In various embodiments thiscontinuum of pre-symptomatic AD can also encompass, but is notnecessarily limited to, (1) individuals who carry one or moreapolipoprotein E (APOE) ε4 alleles who are known or believed to have anincreased risk of developing AD dementia, at the point they are AD-Pbiomarker-positive, and (2) carriers of autosomal dominant mutations,who are in the presymptomatic biomarker-positive stage of their illness,and who will almost certainly manifest clinical symptoms and progress todementia.

A biomarker model has been proposed in which the most widely validatedbiomarkers of AD-P become abnormal and likewise reach a ceiling in anordered manner (see, e.g., Jack et al. (2010) Lancet Neurol., 9:119-128.). This biomarker model parallels proposed pathophysiologicalsequence of (pre-AD/AD), and is relevant to tracking the preclinical(asymptomatic) stages of AD (see, e.g., FIG. 3 in Sperling et al. (2011)Alzheimer's & Dementia, 1-13). Biomarkers of brain amyloidosis include,but are not limited to reductions in CSF Aβ₄₂ and increased amyloidtracer retention on positron emission tomography (PET) imaging. ElevatedCSF tau is not specific to AD and is thought to be a biomarker ofneuronal injury. Decreased fluorodeoxyglucose 18F (FDG) uptake on PETwith a temporoparietal pattern of hypometabolism is a biomarker ofAD-related synaptic dysfunction. Brain atrophy on structural magneticresonance imaging (Mill) in a characteristic pattern involving themedial temporal lobes, paralimbic and temporoparietal cortices is abiomarker of AD-related neurodegeneration. Other markers include, butare not limited to volumetric MM, FDG-PET, or plasma biomarkers (see,e.g., Vemuri et al. (2009) Neurology, 73: 294-301; Yaffe et al. (2011)JAMA 305: 261-266).

In certain embodiments the subjects suitable for the prophylacticmethods contemplated herein include, but are not limited to, subjectscharacterized as having asymptomatic cerebral amyloidosis. In variousembodiments these individuals have biomarker evidence of Aβ accumulationwith elevated tracer retention on PET amyloid imaging and/or low Aβ42 inCSF assay, but typically no detectable evidence of additional brainalterations suggestive of neurodegeneration or subtle cognitive and/orbehavioral symptomatology.

It is noted that currently available CSF and PET imaging biomarkers ofAβ primarily provide evidence of amyloid accumulation and deposition offibrillar forms of amyloid. Data suggest that soluble or oligomericforms of Aβ are likely in equilibrium with plaques, which may serve asreservoirs. In certain embodiments it is contemplated that there is anidentifiable preplaque stage in which only soluble forms of Aβ arepresent. In certain embodiments it is contemplated that oligomeric formsof amyloid may be critical in the pathological cascade, and provideuseful markers. In addition, early synaptic changes may be presentbefore evidence of amyloid accumulation.

In certain embodiments the subjects suitable for the prophylacticmethods contemplated herein include, but are not limited to, subjectscharacterized as amyloid positive with evidence of synaptic dysfunctionand/or early neurodegeneration. In various embodiments these subjectshave evidence of amyloid positivity and presence of one or more markersof “downstream” AD-related neuronal injury. Illustrative, butnon-limiting markers of neuronal injury include, but are not limited to(1) elevated CSF tau or phospho-tau, (2) hypometabolism in an AD-likepattern (i.e., posterior cingulate, precuneus, and/or temporoparietalcortices) on FDG-PET, and (3) cortical thinning/gray matter loss in aspecific anatomic distribution (i.e., lateral and medial parietal,posterior cingulate, and lateral temporal cortices) and/or hippocampalatrophy on volumetric MRI. Other markers include, but are not limited tofMRI measures of default network connectivity. In certain embodimentsearly synaptic dysfunction, as assessed by functional imaging techniquessuch as FDG-PET and fMRI, can be detectable before volumetric loss.Without being bound to a particular theory, it is believed thatamyloid-positive individuals with evidence of early neurodegenerationmay be farther down the trajectory (i.e., in later stages of preclinical(asymptomatic) AD).

In certain embodiments the subjects suitable for the prophylacticmethods contemplated herein include, but are not limited to, subjectscharacterized as amyloid positive with evidence of neurodegeneration andsubtle cognitive decline. Without being bound to a particular theory, itis believed that those individuals with biomarker evidence of amyloidaccumulation, early neurodegeneration, and evidence of subtle cognitivedecline are in the last stage of preclinical (asymptomatic) AD, and areapproaching the border zone with clinical criteria for mild cognitiveimpairment (MCI). These individuals may demonstrate evidence of declinefrom their own baseline (particularly if proxies of cognitive reserveare taken into consideration), even if they still perform within the“normal” range on standard cognitive measures. Without being bound to aparticular theory, it is believed that more sensitive cognitivemeasures, particularly with challenging episodic memory measures, maydetect very subtle cognitive impairment in amyloid-positive individuals.In certain embodiments criteria include, but are not limited to,self-complaint of memory decline or other subtle neurobehavioralchanges.

As indicated above, subjects/patients amenable to prophylactic methodsdescribed herein include individuals at risk of disease (e.g., apathology characterized by amyloid plaque formation such as MCI) but notshowing symptoms, as well as subjects presently showing certain symptomsor markers. It is known that the risk of MCI and later Alzheimer'sdisease generally increases with age. Accordingly, in asymptomaticsubjects with no other known risk factors, in certain embodiments,prophylactic application is contemplated for subjects over 50 years ofage, or subjects over 55 years of age, or subjects over 60 years of age,or subjects over 65 years of age, or subjects over 70 years of age, orsubjects over 75 years of age, or subjects over 80 years of age, inparticular to prevent or slow the onset or ultimate severity of mildcognitive impairment (MCI), and/or to slow or prevent the progressionfrom MCI to early stage Alzheimer's disease (AD).

In certain embodiments, the methods described herein are especiallyuseful for individuals who do have a known genetic risk of Alzheimer'sdisease (or other amyloidogenic pathologies), whether they areasymptomatic or showing symptoms of disease. Such individuals includethose having relatives who have experienced MCI or AD (e.g., a parent, agrandparent, a sibling), and those whose risk is determined by analysisof genetic or biochemical markers. Genetic markers of risk towardAlzheimer's disease include, for example, mutations in the APP gene,particularly mutations at position 717 and positions 670 and 671referred to as the Hardy and Swedish mutations respectively (see Hardy(1997) Trends. Neurosci., 20: 154-159). Other markers of risk includemutations in the presenilin genes (PSI and PS2), family history of AD,having the familial Alzheimer's disease (FAD) mutation, the APOE c4allele, hypercholesterolemia or atherosclerosis. Further susceptibilitygenes for the development of Alzheimer's disease are reviewed, e.g., inSleegers, et al. (2010) Trends Genet. 26(2): 84-93.

In some embodiments, the subject is asymptomatic but has familial and/orgenetic risk factors for developing MCI or Alzheimer's disease. Inasymptomatic patients, treatment can begin at any age (e.g., at about20, about 30, about 40, about 50 years of age). Usually, however, it isnot necessary to begin treatment until a patient reaches at least about40, or at least about 50, or at least about 55, or at least about 60, orat least about 65, or at least about 70 years of age.

In some embodiments, the subject exhibits symptoms, for example, of mildcognitive impairment (MCI) or Alzheimer's disease (AD). Individualspresently suffering from Alzheimer's disease can be recognized fromcharacteristic dementia, as well as the presence of risk factorsdescribed above. In addition, a number of diagnostic tests are availablefor identifying individuals who have AD. These include measurement ofCSF Tau, phospho-tau (pTau), Aβ42 levels and C-terminally cleaved APPfragment (APPneo). Elevated total-Tau (tTau), phospho-Tau (pTau),APPneo, soluble Aβ40, pTau/Aβ42 ratio and tTau/Aβ42 ratio, and decreasedAβ42 levels, Aβ42/Aβ40 ratio, Aβ42/Aβ38 ratio, sAPPα levels, sAPPα/sAPPβratio, sAPPα/Aβ40 ratio, and sAPPα/Aβ42 ratio signify the presence ofAD. In some embodiments, the subject or patient is diagnosed as havingMCI. Increased levels of neural thread protein (NTP) in urine and/orincreased levels of α2-macroglobulin (α2M) and/or complement factor H(CFH) in plasma are also biomarkers of MCI and/or AD (see, e.g., Anoopet al. (2010) Int. J. Alzheimer's Dis. 2010:606802).

In certain embodiments, subjects amenable to treatment may haveage-associated memory impairment (AAMI), or mild cognitive impairment(MCI). The methods described herein are particularly well-suited to theprophylaxis and/or treatment of MCI. In such instances, the methods candelay or prevent the onset of MCI, and or reduce one or more symptomscharacteristic of MCI and/or delay or prevent the progression from MCIto early-, mid- or late-stage Alzheimer's disease or reduce the ultimateseverity of the disease.

Mild Cognitive Impairment (MCI)

Mild cognitive impairment (MCI, also known as incipient dementia, orisolated memory impairment) is a diagnosis given to individuals who havecognitive impairments beyond that expected for their age and education,but that typically do not interfere significantly with their dailyactivities (see, e.g., Petersen et al. (1999) Arch. Neurol. 56(3):303-308). It is considered in many instances to be a boundary ortransitional stage between normal aging and dementia. Although MCI canpresent with a variety of symptoms, when memory loss is the predominantsymptom it is termed “amnestic MCI” and is frequently seen as a riskfactor for Alzheimer's disease (see, e.g., Grundman et al. (2004) Arch.Neurol. 61(1): 59-66; and on the internet aten.wikipedia.org/wiki/Mild_cognitive_impairment-cite_note-Grundman-1).When individuals have impairments in domains other than memory it isoften classified as non-amnestic single- or multiple-domain MCI andthese individuals are believed to be more likely to convert to otherdementias (e.g., dementia with Lewy bodies). There is evidencesuggesting that while amnestic MCI patients may not meet neuropathologiccriteria for Alzheimer's disease, patients may be in a transitionalstage of evolving Alzheimer's disease; patients in this hypothesizedtransitional stage demonstrated diffuse amyloid in the neocortex andfrequent neurofibrillary tangles in the medial temporal lobe (see, e.g.,Petersen et al. (2006) Arch. Neurol. 63(5): 665-72).

The diagnosis of MCI typically involves a comprehensive clinicalassessment including clinical observation, neuroimaging, blood tests andneuropsychological testing. In certain embodiments diagnostic criteriafor MIC include, but are not limited to those described by Albert et al.(2011) Alzheimer's & Dementia. 1-10. As described therein, diagnosticcriteria include (1) core clinical criteria that could be used byhealthcare providers without access to advanced imaging techniques orcerebrospinal fluid analysis, and (2) research criteria that could beused in clinical research settings, including clinical trials. Thesecond set of criteria incorporate the use of biomarkers based onimaging and cerebrospinal fluid measures. The final set of criteria formild cognitive impairment due to AD has four levels of certainty,depending on the presence and nature of the biomarker findings.

In certain embodiments clinical evaluation/diagnosis of MCI involves:(1) Concern reflecting a change in cognition reported by patient orinformant or clinician (i.e., historical or observed evidence of declineover time); (2) Objective evidence of Impairment in one or morecognitive domains, typically including memory (i.e., formal or bedsidetesting to establish level of cognitive function in multiple domains);(3) Preservation of independence in functional abilities; (4) Notdemented; and in certain embodiments, (5) An etiology of MCI consistentwith AD pathophysiological processes. Typically vascular, traumatic, andmedical causes of cognitive decline, are ruled out where possible. Incertain embodiments, when feasible, evidence of longitudinal decline incognition is identified. Diagnosis is reinforced by a history consistentwith AD genetic factors, where relevant.

With respect to impairment in cognitive domain(s), there should beevidence of concern about a change in cognition, in comparison with theperson's previous level. There should be evidence of lower performancein one or more cognitive domains that is greater than would be expectedfor the patient's age and educational background. If repeatedassessments are available, then a decline in performance should beevident over time. This change can occur in a variety of cognitivedomains, including memory, executive function, attention, language, andvisuospatial skills. An impairment in episodic memory (i.e., the abilityto learn and retain new information) is seen most commonly in MCIpatients who subsequently progress to a diagnosis of AD dementia.

With respect to preservation of independence in functional abilities, itis noted that persons with MCI commonly have mild problems performingcomplex functional tasks which they used to perform shopping. They maytake more time, be less efficient, and make more errors at performingsuch activities than in the past. Nevertheless, they generally maintaintheir independence of function in daily life, with minimal aids orassistance.

With respect to dementia, the cognitive changes should be sufficientlymild that there is no evidence of a significant impairment in social oroccupational functioning. If an individual has only been evaluated once,change will be inferred from the history and/or evidence that cognitiveperformance is impaired beyond what would have been expected for thatindividual.

Cognitive testing is optimal for objectively assessing the degree ofcognitive impairment for an individual. Scores on cognitive tests forindividuals with MCI are typically 1 to 1.5 standard deviations belowthe mean for their age and education matched peers on culturallyappropriate normative data (i.e., for the impaired domain(s), whenavailable).

Episodic memory (i.e., the ability to learn and retain new information)is most commonly seen in MCI patients who subsequently progress to adiagnosis of AD dementia. There are a variety of episodic memory teststhat are useful for identifying those MCI patients who have a highlikelihood of progressing to AD dementia within a few years. These teststypically assess both immediate and delayed recall, so that it ispossible to determine retention over a delay. Many, although not all, ofthe tests that have proven useful in this regard are wordlist learningtests with multiple trials. Such tests reveal the rate of learning overtime, as well as the maximum amount acquired over the course of thelearning trials. They are also useful for demonstrating that theindividual is, in fact, paying attention to the task on immediaterecall, which then can be used as a baseline to assess the relativeamount of material retained on delayed recall. Examples of such testsinclude (but are not limited to: the Free and Cued Selective RemindingTest, the Rey Auditory Verbal Learning Test, and the California VerbalLearning Test. Other episodic memory measures include, but are notlimited to: immediate and delayed recall of a paragraph such as theLogical Memory I and II of the Wechsler Memory Scale Revised (or otherversions) and immediate and delayed recall of nonverbal materials, suchas the Visual Reproduction subtests of the Wechsler Memory Scale-RevisedI and II.

Because other cognitive domains can be impaired among individuals withMCI, it is desirable to examine domains in addition to memory. Theseinclude, but are not limited to executive functions (e.g., set-shifting,reasoning, problem-solving, planning), language (e.g., naming, fluency,expressive speech, and comprehension), visuospatial skills, andattentional control (e.g., simple and divided attention). Many clinicalneuropsychological measures are available to assess these cognitivedomains, including (but not limited to the Trail Making Test (executivefunction), the Boston Naming Test, letter and category fluency(language), figure copying (spatial skills), and digit span forward(attention).

As indicated above, genetic factors can be incorporated into thediagnosis of MCI. If an autosomal dominant form of AD is known to bepresent (i.e., mutation in APP, PS1, PS2), then the development of MCIis most likely the precursor to AD dementia. The large majority of thesecases develop early onset AD (i.e., onset below 65 years of age).

In addition, there are genetic influences on the development of lateonset AD dementia. For example, the presence of one or two c4 alleles inthe apolipoprotein E (APOE) gene is a genetic variant broadly acceptedas increasing risk for late-onset AD dementia. Evidence suggests that anindividual who meets the clinical, cognitive, and etiologic criteria forMCI, and is also APOE c4 positive, is more likely to progress to ADdementia within a few years than an individual without this geneticcharacteristic. It is believed that additional genes play an important,but smaller role than APOE and also confer changes in risk forprogression to AD dementia (see, e.g., Bertram et al. (2010) Neuron, 21:270-281).

In certain embodiments subjects suitable for the prophylactic methodsdescribed herein include, but need not be limited to, subjectsidentified having one or more of the core clinical criteria describedabove and/or subjects identified with one or more “research criteria”for MCI, e.g., as described below.

“Research criteria” for the identification/prognosis of MCI include, butare not limited to biomarkers that increase the likelihood that MCIsyndrome is due to the pathophysiological processes of AD. Without beingbound to a particular theory, it is believed that the conjointapplication of clinical criteria and biomarkers can result in variouslevels of certainty that the MCI syndrome is due to ADpathophysiological processes. In certain embodiments, two categories ofbiomarkers have been the most studied and applied to clinical outcomesare contemplated. These include “Aβ” (which includes CSF Aβ₄₂ and/or PETamyloid imaging) and “biomarkers of neuronal injury” (which include, butare not limited to CSF tau/p-tau, hippocampal, or medial temporal lobeatrophy on MRI, and temporoparietal/precuneus hypometabolism orhypoperfusion on PET or SPECT).

Without being bound to a particular theory, it is believed that evidenceof both Aβ, and neuronal injury (either an increase in tau/p-tau orimaging biomarkers in a topographical pattern characteristic of AD),together confers the highest probability that the AD pathophysiologicalprocess is present. Conversely, if these biomarkers are negative, thismay provide information concerning the likelihood of an alternatediagnosis. It is recognized that biomarker findings may be contradictoryand accordingly any biomarker combination is indicative (an indicator)used on the context of a differential diagnosis and not itselfdispositive. It is recognized that varying severities of an abnormalitymay confer different likelihoods or prognoses, that are difficult toquantify accurately for broad application.

For those potential MCI subjects whose clinical and cognitive MCIsyndrome is consistent with AD as the etiology, the addition ofbiomarker analysis effects levels of certainty in the diagnosis. In themost typical example in which the clinical and cognitive syndrome of MCIhas been established, including evidence of an episodic memory disorderand a presumed degenerative etiology, the most likely cause is theneurodegenerative process of AD. However, the eventual outcome still hasvariable degrees of certainty. The likelihood of progression to ADdementia will vary with the severity of the cognitive decline and thenature of the evidence suggesting that AD pathophysiology is theunderlying cause. Without being bound to a particular theory it isbelieved that positive biomarkers reflecting neuronal injury increasethe likelihood that progression to dementia will occur within a fewyears and that positive findings reflecting both Aβ accumulation andneuronal injury together confer the highest likelihood that thediagnosis is MCI due to AD.

A positive Aβ biomarker and a positive biomarker of neuronal injuryprovide an indication that the MCI syndrome is due to AD processes andthe subject is well suited for the methods described herein.

A positive Aβ biomarker in a situation in which neuronal injurybiomarkers have not been or cannot be tested or a positive biomarker ofneuronal injury in a situation in which Aβ biomarkers have not been orcannot be tested indicate an intermediate likelihood that the MCIsyndrome is due to AD. Such subjects are believed to be is well suitedfor the methods described herein

Negative biomarkers for both Aβ and neuronal injury suggest that the MCIsyndrome is not due to AD. In such instances the subjects may not bewell suited for the methods described herein.

There is evidence that magnetic resonance imaging can observedeterioration, including progressive loss of gray matter in the brain,from mild cognitive impairment to full-blown Alzheimer disease (see,e.g., Whitwell et al. (2008) Neurology 70(7): 512-520). A techniqueknown as PiB PET imaging is used to clearly show the sites and shapes ofbeta amyloid deposits in living subjects using a C11 tracer that bindsselectively to such deposits (see, e.g., Jack et al. (2008) Brain 131(Pt3): 665-680).

In certain embodiments, MCI is typically diagnosed when there is 1)Evidence of memory impairment; 2) Preservation of general cognitive andfunctional abilities; and 3) Absence of diagnosed dementia.

In certain embodiments MCI and stages of Alzheimer's disease can beidentified/categorized, in part by Clinical Dementia Rating (CDR)scores. The CDR is a five point scale used to characterize six domainsof cognitive and functional performance applicable to Alzheimer diseaseand related dementias: Memory, Orientation, Judgment & Problem Solving,Community Affairs, Home & Hobbies, and Personal Care. The information tomake each rating can be obtained through a semi-structured interview ofthe patient and a reliable informant or collateral source (e.g., familymember).

The CDR table provides descriptive anchors that guide the clinician inmaking appropriate ratings based on interview data and clinicaljudgment. In addition to ratings for each domain, an overall CDR scoremay be calculated through the use of an algorithm. This score is usefulfor characterizing and tracking a patient's level ofimpairment/dementia: 0=Normal; 0.5=Very Mild Dementia; 1=Mild Dementia;2=Moderate Dementia; and 3=Severe Dementia. An illustrative CDR table isshown in Table 3.

TABLE 3 Illustrative clinical dementia rating (CDR) table. Impairment:None Questionable Mild Moderate Severe CDR: 0 0.5 1 2 3 Memory No memoryConsistent Moderate Severe Severe loss or slight slight memory loss;memory memory inconsistent forgetfulness; more marked loss; only loss;only forgetfulness partial for recent highly fragments recollectionevents; defect learned remain of events' interferes material “benign”with retained; forgetfulness everyday new material activities rapidlylost Orientation Fully Fully Moderate Severe Oriented to orientedoriented difficulty difficulty person only except for with time withtime slight relationships; relationships; difficulty oriented forusually with time place at disoriented relationships examination; totime, often may have to place. geographic disorientation elsewhereJudgment & Solves Slight Moderate Severely Unable to Problem everydayimpairment difficulty in impaired in make Solving problems & in solvinghandling handling judgments handles problems, problems, problems, orsolve business & similarities, similarities similarities problemsfinancial and and and affairs well; differences differences;differences; judgment social social good in judgment judgment relationto usually usually past maintained impaired performance CommunityIndependent Slight Unable to No pretense of independent Affairs functionat impairment function function outside of home usual level in theseindependently Appears well Appears too in job, activities at theseenough to be ill to be shopping, activities taken to taken to volunteer,although may functions functions and social still be outside a outside agroups engaged in family family some; home home. appears normal tocasual inspection Home and Life at Life at home, Mild bit Only simple NoHobbies home, hobbies, and definite chores significant hobbies, andintellectual impairment preserved; function in intellectual interests offunction at very home interests slightly home; more restricted wellimpaired difficult interests, maintained chores poorly abandoned;maintained more complicated hobbies and interests abandoned PersonalFully capable of self-care Needs Requires Requires Care promptingassistance in much help dressing, with hygiene, personal keeping ofcare; personal frequent effects incontinence

A CDR rating of ˜0.5 or ˜0.5 to 1.0 is often considered clinicallyrelevant MCI. Higher CDR ratings can be indicative of progression intoAlzheimer's disease.

In certain embodiments administration of one or more agents describedherein (e.g., triazolopyrimidine(s) and/or triazolopyridine(s) describedherein, or tautomer(s) or stereoisomer(s) thereof, or pharmaceuticallyacceptable salts, solvates, or clathrates of said triazolopyrimidine(s)and/or triazolopyridine(s), said stereoisomer(s), or said tautomer(s),or analogues, derivatives, or prodrugs thereof) is deemed effective whenthere is a reduction in the CSF of levels of one or more componentsselected from the group consisting of Tau, phospho-Tau (pTau), APPneo,soluble Aβ40, soluble Aβ42, and/or Aβ42/Aβ40 ratio, and/or when there isa reduction of the plaque load in the brain of the subject, and/or whenthere is a reduction in the rate of plaque formation in the brain of thesubject, and/or when there is an improvement in the cognitive abilitiesof the subject, and/or when there is a perceived improvement in qualityof life by the subject, and/or when there is a significant reduction inclinical dementia rating (CDR), and/or when the rate of increase inclinical dementia rating is slowed or stopped and/or when theprogression from MCI to early stage AD is slowed or stopped.

In some embodiments, a diagnosis of MCI can be determined by consideringthe results of several clinical tests. For example, Grundman, et al.(2004) Arch. Neurol. 61: 59-66, report that a diagnosis of MCI can beestablished with clinical efficiency using a simple memory test(paragraph recall) to establish an objective memory deficit, a measureof general cognition (Mini-Mental State Exam (MMSE), discussed ingreater detail below) to exclude a broader cognitive decline beyondmemory, and a structured clinical interview (CDR) with patients andcaregivers to verify the patient's memory complaint and memory loss andto ensure that the patient was not demented. Patients with MCI perform,on average, less than 1 standard deviation (SD) below normal onnonmemorycognitive measures included in the battery. Tests of learning,attention, perceptual speed, category fluency, and executive functionmay be impaired in patients with MCI, but these are far less prominentthan the memory deficit.

Alzheimer's Disease (AD).

In certain embodiments the active agent(s) (e.g., triazolopyrimidine(s)and/or triazolopyridine(s) described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts, solvates,or clathrates of said triazolopyrimidine and/or triazolopyridine(s),said stereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof) are contemplated for the treatment of Alzheimer'sdisease. In such instances the methods described herein are useful inpreventing or slowing the onset of Alzheimer's disease (AD), in reducingthe severity of AD when the subject has transitioned to clinical ADdiagnosis, and/or in mitigating one or more symptoms of Alzheimer'sdisease.

In particular, where the Alzheimer's disease is early stage, the methodscan reduce or eliminate one or more symptoms characteristic of AD and/ordelay or prevent the progression from MCI to early or later stageAlzheimer's disease.

Individuals presently suffering from Alzheimer's disease can berecognized from characteristic dementia, as well as the presence of riskfactors described above. In addition, a number of diagnostic tests areavailable for identifying individuals who have AD. Individuals presentlysuffering from Alzheimer's disease can be recognized from characteristicdementia, as well as the presence of risk factors described above. Inaddition, a number of diagnostic tests are available for identifyingindividuals who have AD. These include measurement of CSF Tau,phospho-tau (pTau), sAPPα, sAPPβ, Aβ40, Aβ42 levels and/or C terminallycleaved APP fragment (APPneo). Elevated Tau, pTau, sAPPβ and/or APPneo,and/or decreased sAPPα, soluble Aβ40 and/or soluble Aβ42 levels,particularly in the context of a differential diagnosis, can signify thepresence of AD.

In certain embodiments subjects amenable to treatment may haveAlzheimer's disease. Individuals suffering from Alzheimer's disease canalso be diagnosed by Alzheimer's disease and Related DisordersAssociation (ADRDA) criteria. The NINCDS-ADRDA Alzheimer's Criteria wereproposed in 1984 by the National Institute of Neurological andCommunicative Disorders and Stroke and the Alzheimer's Disease andRelated Disorders Association (now known as the Alzheimer's Association)and are among the most used in the diagnosis of Alzheimer's disease(AD). McKhann, et al. (1984) Neurology 34(7): 939-44. According to thesecriteria, the presence of cognitive impairment and a suspected dementiasyndrome should be confirmed by neuropsychological testing for aclinical diagnosis of possible or probable AD. However, histopathologicconfirmation (microscopic examination of brain tissue) is generally usedfor a dispositive diagnosis. The NINCDS-ADRDA Alzheimer's Criteriaspecify eight cognitive domains that may be impaired in AD: memory,language, perceptual skills, attention, constructive abilities,orientation, problem solving and functional abilities). These criteriahave shown good reliability and validity.

Baseline evaluations of patient function can made using classicpsychometric measures, such as the Mini-Mental State Exam (MMSE)(Folstein et al. (1975) J. Psychiatric Research 12 (3): 189-198), andthe Alzheimer's Disease Assessment Scale (ADAS), which is acomprehensive scale for evaluating patients with Alzheimer's Diseasestatus and function (see, e.g., Rosen, et al. (1984) Am. J. Psychiatr.,141: 1356-1364). These psychometric scales provide a measure ofprogression of the Alzheimer's condition. Suitable qualitative lifescales can also be used to monitor treatment. The extent of diseaseprogression can be determined using a Mini-Mental State Exam (MMSE)(see, e.g., Folstein, et al. supra). Any score greater than or equal to25 points (out of 30) is effectively normal (intact). Below this, scorescan indicate severe (<9 points), moderate (10-20 points) or mild (21-24points) Alzheimer's disease.

Alzheimer's disease can be broken down into various stages including: 1)Moderate cognitive decline (Mild or early-stage Alzheimer's disease), 2)Moderately severe cognitive decline (Moderate or mid-stage Alzheimer'sdisease), 3) Severe cognitive decline (Moderately severe or mid-stageAlzheimer's disease), and 4) Very severe cognitive decline (Severe orlate-stage Alzheimer's disease) as shown in Table 4.

TABLE 4 Illustrative stages of Alzheimer's disease. Moderate CognitiveDecline (Mild or early stage AD) At this stage, a careful medicalinterview detects clear-cut deficiencies in the following areas:Decreased knowledge of recent events. Impaired ability to performchallenging mental arithmetic. For example, to count backward from 100by 7s. Decreased capacity to perform complex tasks, such as marketing,planning dinner for guests, or paying bills and managing finances.Reduced memory of personal history. The affected individual may seemsubdued and withdrawn, especially in socially or mentally challengingsituations. Moderately severe cognitive decline (Moderate or mid-stageAlzheimer's disease) Major gaps in memory and deficits in cognitivefunction emerge. Some assistance with day-to-day activities becomesessential. At this stage, individuals may: Be unable during a medicalinterview to recall such important details as their current address,their telephone number, or the name of the college or high school fromwhich they graduated. Become confused about where they are or about thedate, day of the week or season. Have trouble with less challengingmental arithmetic; for example, counting backward from 40 by 4s or from20 by 2s. Need help choosing proper clothing for the season or theoccasion. Usually retain substantial knowledge about themselves and knowtheir own name and the names of their spouse or children. Usuallyrequire no assistance with eating or using the toilet. Severe cognitivedecline (Moderately severe or mid-stage Alzheimer's disease) Memorydifficulties continue to worsen, significant personality changes mayemerge, and affected individuals need extensive help with dailyactivities. At this stage, individuals may: Lose most awareness ofrecent experiences and events as well as of their surroundings.Recollect their personal history imperfectly, although they generallyrecall their own name. Occasionally forget the name of their spouse orprimary caregiver but generally can distinguish familiar from unfamiliarfaces. Need help getting dressed properly; without supervision, may makesuch errors as putting pajamas over daytime clothes or shoes on wrongfeet. Experience disruption of their normal sleep/waking cycle. Needhelp with handling details of toileting (flushing toilet, wiping anddisposing of tissue properly). Have increasing episodes of urinary orfecal incontinence. Experience significant personality changes andbehavioral symptoms, including suspiciousness and delusions (forexample, believing that their caregiver is an impostor); hallucinations(seeing or hearing things that are not really there); or compulsive,repetitive behaviors such as hand-wringing or tissue shredding. Tend towander and become lost. Very severe cognitive decline (Severe orlate-stage Alzheimer's disease) This is the final stage of the diseasewhen individuals lose the ability to respond to their environment, theability to speak, and, ultimately, the ability to control movement.Frequently individuals lose their capacity for recognizable speech,although words or phrases may occasionally be uttered. Individuals needhelp with eating and toileting and there is general incontinence.Individuals lose the ability to walk without assistance, then theability to sit without support, the ability to smile, and the ability tohold their head up.Reflexes become abnormal and muscles grow rigid.Swallowing is impaired.

In various embodiments administration of one or more agents describedherein to subjects diagnosed with Alzheimer's disease is deemedeffective when the there is a reduction in the CSF of levels of one ormore components selected from the group consisting of Tau, phospho-Tau(pTau), APPneo, soluble Aβ40, soluble Aβ42, and/or and Aβ42/Aβ40 ratio,and/or when there is a reduction of the plaque load in the brain of thesubject, and/or when there is a reduction in the rate of plaqueformation in the brain of the subject, and/or when there is animprovement in the cognitive abilities of the subject, and/or when thereis a perceived improvement in quality of life by the subject, and/orwhen there is a significant reduction in clinical dementia rating (CDR)of the subject, and/or when the rate of increase in clinical dementiarating is slowed or stopped and/or when the progression of AD is slowedor stopped (e.g., when the transition from one stage to another aslisted in Table 3 is slowed or stopped).

In certain embodiments subjects amenable to the present methodsgenerally are free of a neurological disease or disorder other thanAlzheimer's disease. For example, in certain embodiments, the subjectdoes not have and is not at risk of developing a neurological disease ordisorder such as Parkinson's disease, and/or schizophrenia, and/orpsychosis.

Active Agent(s).

The methods described herein are based, in part, on the discovery thatadministration of one or more active agents (e.g., triazolopyrimidine(s)and/or triazolopyridine(s) described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts, solvates,or clathrates of said triazolopyrimidine(s) and/or triazolopyridine(s),said stereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof) are effective to lower tau and/or p-tau, or to preventthe stress-induced (e.g., cortisol-induced) increase in p-tau and finduse in the treatment and/or prophylaxis of diseases characterized byamyloid deposits in the brain, for example, mild cognitive impairment,Alzheimer's disease, macular degeneration, and the like.

In certain embodiments the active agent is a compound (e.g., atriazolopyrimidine and/or a triazolopyridine) as described below. Incertain embodiments the activate agent comprises a compound according tothe Formula I:

or a pharmaceutically acceptable salt, solvate, or clathrate thereof,where R⁰ is present or absent, and when present is selected from thegroup consisting of CHR, NH, O, and NCHR where R is H, alkyl (e.g.,C1-C6 carbon chain), or aryl (e.g., phenyl, substituted phenyl, orheteroaryl);

R² is

and R¹ is

or is selected from the group consisting of a substituted orunsubstituted cyclic or heterocycle selected from the group consistingof pyridine, pyrimidine, naphthalene, quinolone, isoquinoline,cinnoline, phenyl, substituted phenyl, oxazole, furan, pyran, isoxazole,thiazole, thiophene, pyrole, pyrrolidine, pyrazole, and imidazole; or

R¹ is

and R² is

or is selected from the group consisting of a substituted orunsubstituted cyclic or heterocycle selected from the group consistingof pyridine, pyrimidine, naphthalene, quinolone, isoquinoline,cinnoline, phenyl, substituted phenyl, oxazole, furan, pyran, isoxazole,thiazole, thiophene, pyrole, pyrrolidine, pyrazole, and imidazole; R³ isselected from the group consisting of H, CH₃, ethyl, propyl, butyl, CF₃,NH₂, halogen, and CH₂O where R is H, alkyl (e.g., C1-C6 carbon chain),or aryl (e.g., phenyl, substituted phenyl, or heteroaryl). In certainembodiments, the compound is not J03, J04, J05, J06, J07, J08, J09, J10,JH11, J12, J15, and J17. In certain embodiments, the compound is notJ03, J04, J05, J08, and J17.

In certain embodiments, in the compounds above, R¹ is

and the compound is a compound according to Formula II:

In certain embodiments, in the compounds above, R² is

where R⁴, R⁵, and R⁶ are independently selected from the groupconsisting of H, OH, halogen, methyl, and OCH₃, CF₃, ethyl, aryl, SR,SO₂R, NHCOR, and CO₂R, where R is H, alkyl, or aryl (e.g., where saidalkyl is a C1-C6 carbon chain, and said aryl is phenyl, substitutedphenyl, or heteroaryl). In certain embodiments, in compounds of FormulaI, R² is

and the compound is a compound according to Formula III:

and in certain of these compounds, R¹ is

where R⁴, R⁵, and R⁶ are independently selected from the groupconsisting of H, OH, halogen, methyl, and OCH₃.

In certain embodiments, of any of the foregoing compounds, R⁵ is OCH₃,and, in certain of these embodiments, R⁴ is CH₃, R⁴ is CH₃ and R⁶ is H,R⁴ is OCH₃, or R⁴ is OCH₃ and R⁶ is H. In certain embodiments, of any ofthe foregoing compounds, R⁵ is halogen, or R⁵ is F or Cl. In certain ofthese embodiments, particularly where R⁵ is halogen, R⁴ is CH₃, and/orR⁴ is CH₃ and R⁶ is CH₃, or R⁴ is halogen (e.g., F or Cl). In certainembodiments, R⁶ is H, or R⁶ is CH₃.

In certain embodiments, the compound is a compound according to FormulaII, R¹ is the formula shown below or the compound is a compoundaccording to Formula III and R² is the formula shown below:

where R⁷ and R⁸ are independently H, CH₃, OCH₃, and halogen. In certainembodiments, R⁷ is CH₃ and R⁸ is CH₃.

In certain embodiments, the compound is a compound according to FormulaII where R² is

or a compound according to Formula III where R¹ is

In certain embodiments, the compound is a compound according to FormulaII where R² is

or a compound according to Formula III where R¹ is

In certain embodiments in any of the preceding compounds, R⁰ is NH or R⁰is absent.

In certain embodiments, the compound is a compound described in Table 5,or shown in FIG. 2.

TABLE 5 Illustrative, but non-limiting, list of triazolopyrimidineand/or a triazolopyridine compounds. Compound Structure Alsocontemplated J03

J04

J05

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J06

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J07

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J08

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J09

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J10

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J11

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J12

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J14

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J15

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J17

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J19

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J20

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J21

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J22

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J23

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J24

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine J25

Triazolopyridine form Substituents A and B reversed on triazolopyridineor triazolopyrimidine

In certain embodiments the compound comprises a compound selected fromthe group consisting of J06, J07, J11, J12, J14, J15, J19, J20, J21,J22, J23, J24, J25, or a pharmaceutically acceptable salt or solvatethereof. In certain embodiments the compound comprises a compoundselected from the group consisting of J14, J19, J20, J21, J22, J23, J24,and J25, or a pharmaceutically acceptable salt or solvate thereof.

In certain embodiments, the compound described above is effective todecrease in corticotropin-releasing factor (CRF-1) induced p-tau.

In certain embodiments, any of the preceding compounds is provided as aracemic mixture.

In certain embodiments, any of the preceding compounds is asubstantially pure S enantiomer.

In certain embodiments, any of the preceding compounds is asubstantially pure R enantiomer.

It is to be noted (e.g., as indicated above in Table 5) that wherever atriazolopyrimidine is described herein the correspondingtriazolopyridine (having the same substituents) is contemplated.Conversely, wherever a triazolopyridine is described herein thecorresponding triazolopyrimidine (having the same substituents) iscontemplated. Moreover, for any triazolopyrimidine having substituents Aand B, e.g., as illustrated in FIG. 1, a triazolopyridine and atriazolopyrimidine having substituents A and B reversed is contemplated,and for any triazolopyrimidine having substituents A and B, e.g., asillustrated in FIG. 1, a triazolopyridine and a triazolopyrimidinehaving substituents A and B reversed is contemplated. Additionally, withrespect to any of the triazolopyridines and a triazolopyrimidinesdescribed and/or contemplated herein, a racemic mixture of enantiomersis contemplated as well as a substantially pure (R) enantiomer or asubstantially pure (S) enantiomer.

Various illustrative, but non-limiting triazolopyrimidine and/ortriazolopyridines are also shown in FIGS. 2 and 4. In certainembodiments pharmaceutically acceptable salts, solvates, clathrates,tautomers, pharmaceutically acceptable salts of a tautomer, enantiomersthereof, and pharmaceutically acceptable salts of an enantiomer arecontemplated.

Methods of preparing triazolopyrimidine(s) and/or triazolopyridine(s)such as are described herein are known to those of skill in the art.Generally, in one approach, the relevant triazolopyrimidine and/ortriazolopyridine is illustrated in Example 1, below, describing thesynthesis of J19. As illustrated therein, the synthesis of J19 involvesthe preparation of 6-Chloro-N4-substituted pyrimidine-4,5-diaminefollowed by a cyclisation to the triazolopyrimidine and displacement ofthe chlorine on the pyrimidine ring to yield the desired analog. Asimilar synthetic pathway would be used for the other analogs describedherein. Additionally, a similar synthetic pathway would be used for thetriazolopyridine series, the final product would involve separation ofthe pyridine isomers.

One illustrative, but non-limiting, protocol for the synthesis of J19 isprovided in FIG. 3 and Example 1. Synthesis of additional compoundsdescribed herein are straightforward variations of the synthesis schemesprovided herein.

The various active agents and synthesis schemes are intended to beillustrative and not limiting. Using the teachings provided herein,numerous other (e.g., triazolopyrimidine(s) and/or triazolopyridines ora tautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts, solvates, or clathrates of said triazolopyrimidine and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof can be synthesized andidentified by one of skill in the art.

Pharmaceutical Formulations.

In certain embodiments one or more active agents described herein (e.g.,triazolopyrimidine(s) and/or triazolopyridines described herein, or atautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts, solvates, or clathrates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) are administered to amammal in need thereof, e.g., to a mammal at risk for or suffering froma pathology characterized by abnormal processing of amyloid precursorproteins, a mammal at risk for progression of MCI to Alzheimer'sdisease, and so forth. In certain embodiments the active agent(s) areadministered to prevent or delay the onset of a pre-Alzheimer'scondition and/or cognitive dysfunction, and/or to ameliorate one or moresymptoms of a pre-Alzheimer's cognitive dysfunction, and/or to preventor delay the progression of a pre-Alzheimer's condition or cognitivedysfunction to Alzheimer's disease, and/or to promote the processing ofamyloid precursor protein (APP) by a non-amyloidogenic pathway.

In certain embodiments one or more active agents described herein (e.g.,triazolopyrimidine(s) and/or triazolopyridine(s) described herein, or atautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts or solvates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) are administered to amammal in need thereof, e.g., to a mammal at risk for or suffering froma pathology characterized by abnormal processing of amyloid precursorproteins in conditions other than Alzheimer's disease of MCI.Illustrative conditions, include, but are not limited to AD-typesymptoms of patients with Down's syndrome, glaucoma, maculardegeneration (e.g., age-related macular degeneration (AMD), olfactoryimpairment. in the treatment of type-II diabetes, including diabetesassociated with amyloidogenesis., neurodegenerative diseases such asscrapie, bovine spongiform encaphalopathies (e.g., BSE), traumatic braininjury (“TBI”), Creutzfeld-Jakob disease and the like, type II diabetes.Other conditions characterized by characterized by amyloidformation/deposition are contemplated. Such conditions include, but arenot limited to Huntington's Disease, medullary carcinoma of the thyroid,cardiac arrhythmias, isolated atrial amyloidosis, atherosclerosis,rheumatoid arthritis, aortic medial amyloid, prolactinomas, familialamyloid polyneuropathy, hereditary non-neuropathic systemic amyloidosis,dialysis related amyloidosis, Finnish amyloidosis, Lattice cornealdystrophy, cerebral amyloid angiopathy (e.g., Icelandic type), systemicAL amyloidosis, sporadic inclusion body myositis, cerebrovasculardementia, and the like.

The active agent(s) (e.g., triazolopyrimidine(s) and/ortriazolopyridines described herein) can be administered in the “native”form or, if desired, in the form of salts, esters, amides, prodrugs,derivatives, and the like, provided the salt, ester, amide, prodrug orderivative is suitable pharmacologically, i.e., effective in the presentmethod(s). Salts, esters, amides, prodrugs and other derivatives of theactive agents can be prepared using standard procedures known to thoseskilled in the art of synthetic organic chemistry and described, forexample, by March (1992) Advanced Organic Chemistry; Reactions,Mechanisms and Structure, 4th Ed. N.Y. Wiley-Interscience, and asdescribed above.

For example, a pharmaceutically acceptable salt can be prepared for anyof the agent(s) described herein having a functionality capable offorming a salt. A pharmaceutically acceptable salt is any salt thatretains the activity of the parent compound and does not impart anydeleterious or untoward effect on the subject to which it isadministered and in the context in which it is administered.

In various embodiments pharmaceutically acceptable salts may be derivedfrom organic or inorganic bases. The salt may be a mono or polyvalention. Of particular interest are the inorganic ions, lithium, sodium,potassium, calcium, and magnesium. Organic salts may be made withamines, particularly ammonium salts such as mono-, di- and trialkylamines or ethanol amines. Salts may also be formed with caffeine,tromethamine and similar molecules.

Methods of formulating pharmaceutically active agents as salts, esters,amide, prodrugs, and the like are well known to those of skill in theart. For example, salts can be prepared from the free base usingconventional methodology that typically involves reaction with asuitable acid. Generally, the base form of the drug is dissolved in apolar organic solvent such as methanol or ethanol and the acid is addedthereto. The resulting salt either precipitates or can be brought out ofsolution by addition of a less polar solvent. Suitable acids forpreparing acid addition salts include, but are not limited to bothorganic acids, e.g., acetic acid, propionic acid, glycolic acid, pyruvicacid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid,mandelic acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, salicylic acid, and the like, as well asinorganic acids, e.g., hydrochloric acid, hydrobromic acid, sulfuricacid, nitric acid, phosphoric acid, and the like. An acid addition saltcan be reconverted to the free base by treatment with a suitable base.Certain particularly preferred acid addition salts of the active agentsherein include halide salts, such as may be prepared using hydrochloricor hydrobromic acids. Conversely, preparation of basic salts of theactive agents of this invention are prepared in a similar manner using apharmaceutically acceptable base such as sodium hydroxide, potassiumhydroxide, ammonium hydroxide, calcium hydroxide, trimethylamine, or thelike. Particularly preferred basic salts include alkali metal salts,e.g., the sodium salt, and copper salts.

For the preparation of salt forms of basic drugs, the pKa of thecounterion is preferably at least about 2 pH units lower than the pKa ofthe drug. Similarly, for the preparation of salt forms of acidic drugs,the pKa of the counterion is preferably at least about 2 pH units higherthan the pKa of the drug. This permits the counterion to bring thesolution's pH to a level lower than the pH_(max) to reach the saltplateau, at which the solubility of salt prevails over the solubility offree acid or base. The generalized rule of difference in pKa units ofthe ionizable group in the active pharmaceutical ingredient (API) and inthe acid or base is meant to make the proton transfer energeticallyfavorable. When the pKa of the API and counterion are not significantlydifferent, a solid complex may form but may rapidly disproportionate(i.e., break down into the individual entities of drug and counterion)in an aqueous environment.

Preferably, the counterion is a pharmaceutically acceptable counterion.Suitable anionic salt forms include, but are not limited to acetate,benzoate, benzylate, bitartrate, bromide, carbonate, chloride, citrate,edetate, edisylate, estolate, fumarate, gluceptate, gluconate,hydrobromide, hydrochloride, iodide, lactate, lactobionate, malate,maleate, mandelate, mesylate, methyl bromide, methyl sulfate, mucate,napsylate, nitrate, pamoate (embonate), phosphate and diphosphate,salicylate and disalicylate, stearate, succinate, sulfate, tartrate,tosylate, triethiodide, valerate, and the like, while suitable cationicsalt forms include, but are not limited to aluminum, benzathine,calcium, ethylene diamine, lysine, magnesium, meglumine, potassium,procaine, sodium, tromethamine, zinc, and the like.

Preparation of esters typically involves functionalization of hydroxyland/or carboxyl groups that are present within the molecular structureof the active agent. In certain embodiments, the esters are typicallyacyl-substituted derivatives of free alcohol groups, i.e., moieties thatare derived from carboxylic acids of the formula RCOOH where R is alky,and preferably is lower alkyl. Esters can be reconverted to the freeacids, if desired, by using conventional hydrogenolysis or hydrolysisprocedures.

Amides can also be prepared using techniques known to those skilled inthe art or described in the pertinent literature. For example, amidesmay be prepared from esters, using suitable amine reactants, or they maybe prepared from an anhydride or an acid chloride by reaction withammonia or a lower alkyl amine.

In various embodiments, the active agents identified herein (e.g.,triazolopyrimidine(s) and/or triazolopyridines described herein, or atautomer or stereoisomer thereof, or pharmaceutically acceptable saltsor solvates of said triazolopyrimidine(s) and/or triazolopyridine(s),said stereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof) are useful for parenteral administration, topicaladministration, oral administration, nasal administration (or otherwiseinhaled), rectal administration, or local administration, such as byaerosol or transdermally, for prophylactic and/or therapeutic treatmentof one or more of the pathologies/indications described herein (e.g.,pathologies characterized by excess amyloid plaque formation and/ordeposition or undesired amyloid or pre-amyloid processing).

In various embodiments the active agents described herein can also becombined with a pharmaceutically acceptable carrier (excipient) to forma pharmacological composition. Pharmaceutically acceptable carriers cancontain one or more physiologically acceptable compound(s) that act, forexample, to stabilize the composition or to increase or decrease theabsorption of the active agent(s). Physiologically acceptable compoundscan include, for example, carbohydrates, such as glucose, sucrose, ordextrans, antioxidants, such as ascorbic acid or glutathione, chelatingagents, low molecular weight proteins, protection and uptake enhancerssuch as lipids, compositions that reduce the clearance or hydrolysis ofthe active agents, or excipients or other stabilizers and/or buffers.

Other physiologically acceptable compounds, particularly of use in thepreparation of tablets, capsules, gel caps, and the like include, butare not limited to binders, diluent/fillers, disintegrants, lubricants,suspending agents, and the like.

In certain embodiments, to manufacture an oral dosage form (e.g., atablet), an excipient (e.g., lactose, sucrose, starch, mannitol, etc.),an optional disintegrator (e.g. calcium carbonate,carboxymethylcellulose calcium, sodium starch glycollate, crospovidoneetc.), a binder (e.g. alpha-starch, gum arabic, microcrystallinecellulose, carboxymethylcellulose, polyvinylpyrrolidone,hydroxypropylcellulose, cyclodextrin, etc.), and an optional lubricant(e.g., talc, magnesium stearate, polyethylene glycol 6000, etc.), forinstance, are added to the active component or components (e.g.,triazolopyrimidine(s) and/or triazolopyridine(s) described herein, or atautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts, solvates, or clathrates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) and the resultingcomposition is compressed. Where necessary the compressed product iscoated, e.g., using known methods for masking the taste or for entericdissolution or sustained release. Suitable coating materials include,but are not limited to ethyl-cellulose, hydroxymethylcellulose,POLYOX®yethylene glycol, cellulose acetate phthalate,hydroxypropylmethylcellulose phthalate, and Eudragit (Rohm & Haas,Germany; methacrylic-acrylic copolymer).

Other physiologically acceptable compounds include wetting agents,emulsifying agents, dispersing agents or preservatives that areparticularly useful for preventing the growth or action ofmicroorganisms. Various preservatives are well known and include, forexample, phenol and ascorbic acid. One skilled in the art wouldappreciate that the choice of pharmaceutically acceptable carrier(s),including a physiologically acceptable compound depends, for example, onthe route of administration of the active agent(s) and on the particularphysiochemical characteristics of the active agent(s).

In certain embodiments the excipients are sterile and generally free ofundesirable matter. These compositions can be sterilized byconventional, well-known sterilization techniques. For various oraldosage form excipients such as tablets and capsules sterility is notrequired. The USP/NF standard is usually sufficient.

The pharmaceutical compositions can be administered in a variety of unitdosage forms depending upon the method of administration. Suitable unitdosage forms, include, but are not limited to powders, tablets, pills,capsules, lozenges, suppositories, patches, nasal sprays, injectibles,implantable sustained-release formulations, mucoadherent films, topicalvarnishes, lipid complexes, etc.

Pharmaceutical compositions comprising the active agents describedherein (e.g., triazolopyrimidine(s) and/or triazolopyridine(s) describedherein, or a tautomer(s) or stereoisomer(s) thereof, or pharmaceuticallyacceptable salts or solvates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) can be manufactured bymeans of conventional mixing, dissolving, granulating, dragee-making,levigating, emulsifying, encapsulating, entrapping or lyophilizingprocesses. Pharmaceutical compositions can be formulated in aconventional manner using one or more physiologically acceptablecarriers, diluents, excipients or auxiliaries that facilitate processingof the active agent(s) into preparations that can be usedpharmaceutically. Proper formulation is dependent upon the route ofadministration chosen.

In certain embodiments, the active agents described herein areformulated for oral administration. For oral administration, suitableformulations can be readily formulated by combining the active agent(s)with pharmaceutically acceptable carriers suitable for oral deliverywell known in the art. Such carriers enable the active agent(s)described herein to be formulated as tablets, pills, dragees, caplets,lizenges, gelcaps, capsules, liquids, gels, syrups, slurries,suspensions and the like, for oral ingestion by a patient to be treated.For oral solid formulations such as, for example, powders, capsules andtablets, suitable excipients can include fillers such as sugars (e.g.,lactose, sucrose, mannitol and sorbitol), cellulose preparations (e.g.,maize starch, wheat starch, rice starch, potato starch, gelatin, gumtragacanth, methyl cellulose, hydroxypropylmethyl-cellulose, sodiumcarboxymethylcellulose), synthetic polymers (e.g., polyvinylpyrrolidone(PVP)), granulating agents; and binding agents. If desired,disintegrating agents may be added, such as the cross-linkedpolyvinylpyrrolidone, agar, or alginic acid or a salt thereof such assodium alginate. If desired, solid dosage forms may be sugar-coated orenteric-coated using standard techniques. The preparation ofenteric-coated particles is disclosed for example in U.S. Pat. Nos.4,786,505 and 4,853,230.

For administration by inhalation, the active agent(s) are convenientlydelivered in the form of an aerosol spray from pressurized packs or anebulizer, with the use of a suitable propellant, e.g.,dichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol the dosage unit can be determined byproviding a valve to deliver a metered amount. Capsules and cartridgesof e.g. gelatin for use in an inhaler or insufflator may be formulatedcontaining a powder mix of the compound and a suitable powder base suchas lactose or starch.

In various embodiments the active agent(s) can be formulated in rectalor vaginal compositions such as suppositories or retention enemas, e.g.,containing conventional suppository bases such as cocoa butter or otherglycerides. Methods of formulating active agents for rectal or vaginaldelivery are well known to those of skill in the art (see, e.g., Allen(2007) Suppositories, Pharmaceutical Press) and typically involvecombining the active agents with a suitable base (e.g., hydrophilic(PEG), lipophilic materials such as cocoa butter or Witepsol W45,amphiphilic materials such as Suppocire AP and polyglycolized glyceride,and the like). The base is selected and compounded for a desiredmelting/delivery profile.

For topical administration the active agent(s) described herein (e.g.,triazolopyrimidine(s) and/or triazolopyridine(s) described herein, or atautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts or solvates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) can be formulated assolutions, gels, ointments, creams, suspensions, and the like as arewell-known in the art.

In certain embodiments the active agents described herein are formulatedfor systemic administration (e.g., as an injectable) in accordance withstandard methods well known to those of skill in the art. Systemicformulations include, but are not limited to, those designed foradministration by injection, e.g. subcutaneous, intravenous,intramuscular, intrathecal or intraperitoneal injection, as well asthose designed for transdermal, transmucosal oral or pulmonaryadministration. For injection, the active agents described herein can beformulated in aqueous solutions, preferably in physiologicallycompatible buffers such as Hanks solution, Ringer's solution, orphysiological saline buffer and/or in certain emulsion formulations. Thesolution(s) can contain formulatory agents such as suspending,stabilizing and/or dispersing agents. In certain embodiments the activeagent(s) can be provided in powder form for constitution with a suitablevehicle, e.g., sterile pyrogen-free water, before use. For transmucosaladministration, and/or for blood/brain barrier passage, penetrantsappropriate to the barrier to be permeated can be used in theformulation. Such penetrants are generally known in the art. Injectableformulations and inhalable formulations are generally provided as asterile or substantially sterile formulation.

In addition to the formulations described previously, the activeagent(s) may also be formulated as a depot preparations. Such longacting formulations can be administered by implantation (for examplesubcutaneously or intramuscularly) or by intramuscular injection. Thus,for example, the active agent(s) may be formulated with suitablepolymeric or hydrophobic materials (for example as an emulsion in anacceptable oil) or ion exchange resins, or as sparingly solublederivatives, for example, as a sparingly soluble salt.

In certain embodiments the active agent(s) described herein can also bedelivered through the skin using conventional transdermal drug deliverysystems, i.e., transdermal “patches” wherein the active agent(s) aretypically contained within a laminated structure that serves as a drugdelivery device to be affixed to the skin. In such a structure, the drugcomposition is typically contained in a layer, or “reservoir,”underlying an upper backing layer. It will be appreciated that the term“reservoir” in this context refers to a quantity of “activeingredient(s)” that is ultimately available for delivery to the surfaceof the skin. Thus, for example, the “reservoir” may include the activeingredient(s) in an adhesive on a backing layer of the patch, or in anyof a variety of different matrix formulations known to those of skill inthe art. The patch may contain a single reservoir, or it may containmultiple reservoirs.

In one illustrative embodiment, the reservoir comprises a polymericmatrix of a pharmaceutically acceptable contact adhesive material thatserves to affix the system to the skin during drug delivery. Examples ofsuitable skin contact adhesive materials include, but are not limitedto, polyethylenes, polysiloxanes, polyisobutylenes, polyacrylates,polyurethanes, and the like. Alternatively, the drug-containingreservoir and skin contact adhesive are present as separate and distinctlayers, with the adhesive underlying the reservoir which, in this case,may be either a polymeric matrix as described above, or it may be aliquid or hydrogel reservoir, or may take some other form. The backinglayer in these laminates, which serves as the upper surface of thedevice, preferably functions as a primary structural element of the“patch” and provides the device with much of its flexibility. Thematerial selected for the backing layer is preferably substantiallyimpermeable to the active agent(s) and any other materials that arepresent.

Alternatively, other pharmaceutical delivery systems can be employed.For example, liposomes, emulsions, and microemulsions/nanoemulsions arewell known examples of delivery vehicles that may be used to protect anddeliver pharmaceutically active compounds. Certain organic solvents suchas dimethylsulfoxide also can be employed, although usually at the costof greater toxicity.

In certain embodiments the active agent(s) described herein (e.g.,triazolopyrimidine(s) and/or triazolopyridine(s) described herein, or atautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts, solvates or clathrates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) are formulated in ananoemulsion. Nanoemulsions include, but are not limited to oil in water(O/W) nanoemulsions, and water in oil (W/O) nanoemulsions. Nanoemulsionscan be defined as emulsions with mean droplet diameters ranging fromabout 20 to about 1000 nm. Usually, the average droplet size is betweenabout 20 nm or 50 nm and about 500 nm. The terms sub-micron emulsion(SME) and mini-emulsion are used as synonyms.

Illustrative oil in water (O/W) nanoemulsions include, but are notlimited to: Surfactant micelles—micelles composed of small moleculessurfactants or detergents (e.g., SDS/PBS/2-propanol); Polymermicelles—micelles composed of polymer, copolymer, or block copolymersurfactants (e.g., Pluronic L64/PBS/2-propanol); Blendedmicelles—micelles in which there is more than one surfactant componentor in which one of the liquid phases (generally an alcohol or fatty acidcompound) participates in the formation of the micelle (e.g., octanoicacid/PBS/EtOH); Integral micelles—blended micelles in which the activeagent(s) serve as an auxiliary surfactant, forming an integral part ofthe micelle; and Pickering (solid phase) emulsions—emulsions in whichthe active agent(s) are associated with the exterior of a solidnanoparticle (e.g., polystyrene nanoparticles/PBS/no oil phase).

Illustrative water in oil (W/O) nanoemulsions include, but are notlimited to: Surfactant micelles—micelles composed of small moleculessurfactants or detergents (e.g., dioctyl sulfosuccinate/PBS/2-propanol,isopropylmyristate/PBS/2-propanol, etc.); Polymer micelles—micellescomposed of polymer, copolymer, or block copolymer surfactants (e.g.,PLURONIC® L121/PBS/2-propanol); Blended micelles—micelles in which thereis more than one surfactant component or in which one of the liquidphases (generally an alcohol or fatty acid compound) participates in theformation of the micelle (e.g., capric/caprylic diglyceride/PBS/EtOH);Integral micelles—blended micelles in which the active agent(s) serve asan auxiliary surfactant, forming an integral part of the micelle (e.g.,active agent/PBS/polypropylene glycol); and Pickering (solid phase)emulsions—emulsions in which the active agent(s) are associated with theexterior of a solid nanoparticle (e.g., chitosan nanoparticles/noaqueous phase/mineral oil).

As indicated above, in certain embodiments the nanoemulsions compriseone or more surfactants or detergents. In some embodiments thesurfactant is a non-anionic detergent (e.g., a polysorbate surfactant, apolyoxyethylene ether, etc.). Surfactants that find use in the presentinvention include, but are not limited to surfactants such as theTWEEN®, TRITON®, and TYLOXAPOL® families of compounds.

In certain embodiments the emulsions further comprise one or morecationic halogen containing compounds, including but not limited to,cetylpyridinium chloride. In still further embodiments, the compositionsfurther comprise one or more compounds that increase the interaction(“interaction enhancers”) of the composition with microorganisms (e.g.,chelating agents like ethylenediaminetetraacetic acid, orethylenebis(oxyethylenenitrilo)tetraacetic acid in a buffer).

In some embodiments, the nanoemulsion further comprises an emulsifyingagent to aid in the formation of the emulsion. Emulsifying agentsinclude compounds that aggregate at the oil/water interface to form akind of continuous membrane that prevents direct contact between twoadjacent droplets. Certain embodiments of the present invention featureoil-in-water emulsion compositions that may readily be diluted withwater to a desired concentration without impairing their anti-pathogenicproperties.

In addition to discrete oil droplets dispersed in an aqueous phase,certain oil-in-water emulsions can also contain other lipid structures,such as small lipid vesicles (e.g., lipid spheres that often consist ofseveral substantially concentric lipid bilayers separated from eachother by layers of aqueous phase), micelles (e.g., amphiphilic moleculesin small clusters of 50-200 molecules arranged so that the polar headgroups face outward toward the aqueous phase and the apolar tails aresequestered inward away from the aqueous phase), or lamellar phases(lipid dispersions in which each particle consists of parallelamphiphilic bilayers separated by thin films of water).

These lipid structures are formed as a result of hydrophobic forces thatdrive apolar residues (e.g., long hydrocarbon chains) away from water.The above lipid preparations can generally be described as surfactantlipid preparations (SLPs). SLPs are minimally toxic to mucous membranesand are believed to be metabolized within the small intestine (see e.g.,Hamouda et al. (1998) J Infect. Disease 180: 1939).

In certain embodiments the emulsion comprises a discontinuous oil phasedistributed in an aqueous phase, a first component comprising an alcoholand/or glycerol, and a second component comprising a surfactant or ahalogen-containing compound. The aqueous phase can comprise any type ofaqueous phase including, but not limited to, water (e.g., dionizedwater, distilled water, tap water) and solutions (e.g., phosphatebuffered saline solution or other buffer systems). The oil phase cancomprise any type of oil including, but not limited to, plant oils(e.g., soybean oil, avocado oil, flaxseed oil, coconut oil, cottonseedoil, squalene oil, olive oil, canola oil, corn oil, rapeseed oil,safflower oil, and sunflower oil), animal oils (e.g., fish oil), flavoroil, water insoluble vitamins, mineral oil, and motor oil. In certainembodiments, the oil phase comprises 30-90 vol % of the oil-in-wateremulsion (e.g., constitutes 30-90% of the total volume of the finalemulsion), more preferably 50-80%. The formulations need not be limitedto particular surfactants, however in certain embodiments, thesurfactant is a polysorbate surfactant (e.g., TWEEN 20®, TWEEN 40®,TWEEN 60®, and TWEEN 80®), a pheoxypolyethoxyethanol (e.g., TRITON®X-100, X-301, X-165, X-102, and X-200, and TYLOXAPOL®), or sodiumdodecyl sulfate, and the like.

In certain embodiments a halogen-containing component is present. thenature of the halogen-containing compound, in some embodiments thehalogen-containing compound comprises a chloride salt (e.g., NaCl, KCl,etc.), a cetylpyridinium halide, a cetyltrimethylammonium halide, acetyldimethylethylammonium halide, a cetyldimethylbenzylammonium halide,a cetyltributylphosphonium halide, dodecyltrimethylammonium halides,tetradecyltrimethylammonium halides, cetylpyridinium chloride,cetyltrimethylammonium chloride, cetylbenzyldimethylammonium chloride,cetylpyridinium bromide, cetyltrimethylammonium bromide,cetyldimethylethylammonium bromide, cetyltributylphosphonium bromide,dodecyltrimethylammonium bromide, tetradecyltrimethylammonium bromide,and the like

In certain embodiments the emulsion comprises a quaternary ammoniumcompound. Quaternary ammonium compounds include, but are not limited to,N-alkyldimethyl benzyl ammonium saccharinate,1,3,5-Triazine-1,3,5(2H,4H,6H)-triethanol; 1-Decanaminium,N-decyl-N,N-dimethyl-, chloride (or) Didecyl dimethyl ammonium chloride;2-(2-(p-(Diisobuyl)cresosxy)ethoxy)ethyl dimethyl benzyl ammoniumchloride; 2-(2-(p-(Diisobutyl)phenoxy)ethoxy)ethyl dimethyl benzylammonium chloride; alkyl 1 or 3 benzyl-1-(2-hydroxethyl)-2-imidazoliniumchloride; alkyl bis(2-hydroxyethyl)benzyl ammonium chloride; alkyldimethyl benzyl ammonium chloride; alkyl dimethyl 3,4-dichlorobenzylammonium chloride (100% C12); alkyl dimethyl 3,4-dichlorobenzyl ammoniumchloride (50% C14, 40% C12, 10% C16); alkyl dimethyl 3,4-dichlorobenzylammonium chloride (55% C14, 23% C12, 20% C16); alkyl dimethyl benzylammonium chloride; alkyl dimethyl benzyl ammonium chloride (100% C14);alkyl dimethyl benzyl ammonium chloride (100% C16); alkyl dimethylbenzyl ammonium chloride (41% C14, 28% C12); alkyl dimethyl benzylammonium chloride (47% C12, 18% C14); alkyl dimethyl benzyl ammoniumchloride (55% C16, 20% C14); alkyl dimethyl benzyl ammonium chloride(58% C14, 28% C16); alkyl dimethyl benzyl ammonium chloride (60% C14,25% C12); alkyl dimethyl benzyl ammonium chloride (61% C11, 23% C14);alkyl dimethyl benzyl ammonium chloride (61% C12, 23% C14); alkyldimethyl benzyl ammonium chloride (65% C12, 25% C14); alkyl dimethylbenzyl ammonium chloride (67% C12, 24% C14); alkyl dimethyl benzylammonium chloride (67% C12, 25% C14); alkyl dimethyl benzyl ammoniumchloride (90% C14, 5% C12); alkyl dimethyl benzyl ammonium chloride (93%C14, 4% C12); alkyl dimethyl benzyl ammonium chloride (95% C16, 5% C18);alkyl dimethyl benzyl ammonium chloride (and) didecyl dimethyl ammoniumchloride; alkyl dimethyl benzyl ammonium chloride (as in fatty acids);alkyl dimethyl benzyl ammonium chloride (C12-C16); alkyl dimethyl benzylammonium chloride (C12-C18); alkyl dimethyl benzyl and dialkyl dimethylammonium chloride; alkyl dimethyl dimethybenzyl ammonium chloride; alkyldimethyl ethyl ammonium bromide (90% C14, 5% C16, 5% C12); alkyldimethyl ethyl ammonium bromide (mixed alkyl and alkenyl groups as inthe fatty acids of soybean oil); alkyl dimethyl ethylbenzyl ammoniumchloride; alkyl dimethyl ethylbenzyl ammonium chloride (60% C14); alkyldimethyl isoproylbenzyl ammonium chloride (50% C12, 30% C14, 17% C16, 3%C18); alkyl trimethyl ammonium chloride (58% C18, 40% C16, 1% C14, 1%C12); alkyl trimethyl ammonium chloride (90% C18, 10% C16);alkyldimethyl(ethylbenzyl) ammonium chloride (C12-18); Di-(C8-10)-alkyldimethyl ammonium chlorides; dialkyl dimethyl ammonium chloride; dialkyldimethyl ammonium chloride; dialkyl dimethyl ammonium chloride; dialkylmethyl benzyl ammonium chloride; didecyl dimethyl ammonium chloride;diisodecyl dimethyl ammonium chloride; dioctyl dimethyl ammoniumchloride; dodecyl bis(2-hydroxyethyl) octyl hydrogen ammonium chloride;dodecyl dimethyl benzyl ammonium chloride; dodecylcarbamoyl methyldimethyl benzyl ammonium chloride; heptadecyl hydroxyethylimidazoliniumchloride; hexahydro-1,3,5-thris(2-hydroxyethyl)-s-triazine;myristalkonium chloride (and) Quat RNIUM 14;N,N-Dimethyl-2-hydroxypropylammonium chloride polymer; n-alkyl dimethylbenzyl ammonium chloride; n-alkyl dimethyl ethylbenzyl ammoniumchloride; n-tetradecyl dimethyl benzyl ammonium chloride monohydrate;octyl decyl dimethyl ammonium chloride; octyl dodecyl dimethyl ammoniumchloride; octyphenoxyethoxyethyl dimethyl benzyl ammonium chloride;oxydiethylenebis (alkyl dimethyl ammonium chloride); quaternary ammoniumcompounds, dicoco alkyldimethyl, chloride; trimethoxysily propyldimethyl octadecyl ammonium chloride; trimethoxysilyl quats, trimethyldodecylbenzyl ammonium chloride; n-dodecyl dimethyl ethylbenzyl ammoniumchloride; n-hexadecyl dimethyl benzyl ammonium chloride; n-tetradecyldimethyl benzyl ammonium chloride; n-tetradecyl dimethyl ethylbenzylammonium chloride; and n-octadecyl dimethyl benzyl ammonium chloride.

Nanoemulsion formulations and methods of making such are well known tothose of skill in the art and described for example in U.S. Pat. Nos.7,476,393, 7,468,402, 7,314,624, 6,998,426, 6,902,737, 6,689,371,6,541,018, 6,464,990, 6,461,625, 6,419,946, 6,413,527, 6,375,960,6,335,022, 6,274,150, 6,120,778, 6,039,936, 5,925,341, 5,753,241,5,698,219, and 5,152,923 and in Fanun et al. (2009) Microemulsions:Properties and Applications (Surfactant Science), CRC Press, Boca RatanFla.

In certain embodiments, one or more active agents described herein canbe provided as a “concentrate”, e.g., in a storage container (e.g., in apremeasured volume) ready for dilution, or in a soluble capsule readyfor addition to a volume of water, alcohol, hydrogen peroxide, or otherdiluent.

Extended Release (Sustained Release) Formulations.

In certain embodiments “extended release” formulations of the activeagent(s) described herein (e.g., triazolopyrimidine(s) and/ortriazolopyridine(s) described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts orsolvates of said triazolopyrimidine(s) and/or triazolopyridine(s), saidstereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof) are contemplated. In various embodiments such extendedrelease formulations are designed to avoid the high peak plasma levelsof intravenous and conventional immediate release oral dosage forms.

Illustrative sustained-release formulations include, for example,semipermeable matrices of solid polymers containing the therapeuticagent. Various uses of sustained-release materials have been establishedand are well known by those skilled in the art. Sustained-releasecapsules may, depending on their chemical nature, release the compoundsfor a few weeks up to over 100 days. Depending on the chemical natureand the biological stability of the therapeutic reagent, additionalstrategies for stabilization can be employed.

In certain embodiments such “extended release” formulations utilize themucosa and can independently control tablet disintegration (or erosion)and/or drug dissolution and release from the tablet over time to providea safer delivery profile. In certain embodiments the oral formulationsof active agent(s) described herein (e.g., triazolopyrimidine(s) and/ortriazolopyridine(s) described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts orsolvates of said triazolopyrimidine(s) and/or triazolopyridine(s), saidstereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof) provide individual, repetitive doses that include adefined amount of the active agent that is delivered over a definedamount of time.

One illustrative sustained release formulation is a substantiallyhomogeneous composition that comprises about 0.01% to about 99% w/w, orabout 0.1% to about 95%, or about 0.1%, or about 1%, or about 2%, orabout 5%, or about 10%, or about 15%, or about 20% to about 80%, or toabout 90%, or to about 95%, or to about 97%, or to about 98%, or toabout 99%1 of the active ingredient(s) (e.g., triazolopyrimidine(s)and/or triazolopyridine(s) described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts orsolvates of said triazolopyrimidine(s) and/or triazolopyridine(s), saidstereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof) and one or more mucoadhesives (also referred to hereinas “bioadhesives”) that provide for adherence to the targeted mucosa ofthe subject (patient) and that may further comprise one or more of thefollowing: one or more binders that provide binding of the excipients ina single tablet; one or more hydrogel forming excipients; one or morebulking agents; one or more lubricants; one or more glidants; one ormore solubilizers; one or more surfactants; one or more flavors; one ormore disintegrants; one or more buffering excipients; one or morecoatings; one or more controlled release modifiers; and one or moreother excipients and factors that modify and control the drug'sdissolution or disintegration time and kinetics or protect the activedrug from degradation.

In various embodiments a sustained release pharmaceutical dosage formfor oral transmucosal delivery can be solid or non-solid. In oneillustrative embodiment, the dosage form is a solid that turns into ahydrogel following contact with saliva.

Suitable excipients include, but are not limited to substances added tothe formulations that are required to produce a commercial product andcan include, but are not limited to: bulking agents, binders,surfactants, bioadhesives, lubricants, disintegrants, stabilizers,solubilizers, glidants, and additives or factors that affect dissolutionor disintegration time. Suitable excipients are not limited to thoseabove, and other suitable nontoxic pharmaceutically acceptable carriersfor use in oral formulations can be found in Remington's PharmaceuticalSciences, 17th Edition, 1985.

In certain embodiments extended release formulations of the activeagent(s) described herein for oral transmucosal drug delivery include atleast one bioadhesive (mucoadhesive) agent or a mixture of severalbioadhesives to promote adhesion to the oral mucosa during drugdelivery. In addition the bioadhesive agents may also be effective incontrolling the dosage form erosion time and/or, the drug dissolutionkinetics over time when the dosage form is wetted. Such mucoadhesivedrug delivery systems are very beneficial, since they can prolong theresidence time of the drug at the site of absorption and increase drugbioavailability. The mucoadhesive polymers forming hydrogels aretypically hydrophilic and swellable, containing numerous hydrogenbond-forming groups, like hydroxyl, carboxyl or amine, which favoradhesion. When used in a dry form, they attract water from the mucosalsurface and swell, leading to polymer/mucus interaction through hydrogenbonding, electrostatic, hydrophobic or van der Waals interaction.

Illustrative suitable mucoadhesive or bioadhesive materials, include,but are not limited to natural, synthetic or biological polymers,lipids, phospholipids, and the like. Examples of natural and/orsynthetic polymers include cellulosic derivatives (such asmethylcellulose, carboxymethyl cellulose, hydroxyethyl cellulose,hydroxyethylmethyl cellulose, etc), natural gums (such as guar gum,xanthan gum, locust bean gum, karaya gum, veegum etc.), polyacrylates(such as CARBOPOL®, polycarbophil, etc), alginates, thiol-containingpolymers, POLYOX®yethylenes, polyethylene glycols (PEG) of all molecularweights (preferably between 1000 and 40,000 Da, of any chemistry, linearor branched), dextrans of all molecular weights (preferably between 1000and 40,000 Da of any source), block copolymers, such as those preparedby combinations of lactic and glycolic acid (PLA, PGA, PLGA of variousviscosities, molecular weights and lactic-to-glycolic acid ratios)polyethylene glycol-polypropylene glycol block copolymers of any numberand combination of repeating units (such as PLURONICS®, TEKTRONIX® orGENAPOL® block copolymers), combination of the above copolymers eitherphysically or chemically linked units (for example PEG-PLA or PEG-PLGAcopolymers) mixtures. Preferably the bioadhesive excipient is selectedfrom the group of polyethylene glycols, POLYOX®yethylenes, polyacrylicacid polymers, such as CARBOPOL® (such as CARBOPOL® 71G, 934P, 971P,974P, and the like) and polycarbophils (such as NOVEON® AA-1, NOVEON®CA-1, NOVEON® CA-2, and the like), cellulose and its derivatives andmost preferably it is polyethylene glycol, carbopol, and/or a cellulosicderivative or a combination thereof.

In certain embodiments the mucoadhesive/bioadhesive excipient istypically present at 1-50% w/w, preferably 1-40% w/w or most preferablybetween 5-30% w/w. A particular formulation may contain one or moredifferent bioadhesives in any combination.

In certain embodiments the formulations for oral transmucosal drugdelivery also include a binder or mixture of two or more binders whichfacilitate binding of the excipients into a single dosage form.Illustrative binders include, binders selected from the group consistingof cellulosic derivatives (such as methylcellulose, carboxymethylcellulose, hydroxyethyl cellulose, hydroxyethylmethyl cellulose, etc.),polyacrylates (such as CARBOPOL®, polycarbophil, etc.), POVIDONE® (allgrades), POLYOX®® of any molecular weight or grade, irradiated or not,starch, polyvinylpyrrolidone (PVP), AVICEL®, and the like. In certainembodiments the binder is typically present at 0.5-60% w/w, preferably1-30% w/w and most preferably 1.5-15% w/w.

In certain embodiments the formulations also include at least onehydrogel-forming excipient. Illustrative hydrogel forming excipientsinclude, but are not limited to those selected from the group consistingof polyethylene glycols and other polymers having an ethylene glycolbackbone, whether homopolymers or cross linked heteropolymers, blockcopolymers using ethylene glycol units, such as POLYOX®yethylenehomopolymers (such as POLYOX®® N10/MW=100,000 POLYOX®-80/MW=200,000;POLYOX® 1105/MW=900,000; POLYOX®-301/MW=4,000,000;POLYOX®-303/MW=7,000,000, POLYOX® WSR-N-60K, all of which are tradenamesof Union Carbide), hydroxypropylmethylcellylose (HPMC) of all molecularweights and grades (such as METOLOSE® 90SH50000, METOLOSE® 90SH30000,all of which are tradenames of Shin-Etsu Chemical company), Poloxamers(such as LUTROL® F-68, LUTROL® F-127, F-105 etc., all tradenames of BASFChemicals), GENAPOL®, polyethylene glycols (PEG, such as PEG-1500,PEG-3500, PEG-4000, PEG-6000, PEG-8000, PEG-12000, PEG-20,000, etc.),natural gums (xanthan gum, locust bean gum, etc.) and cellulosederivatives (HC, HMC, HMPC, HPC, CP, CMC), polyacrylic acid-basedpolymers either as free or cross-linked and combinations thereof,biodegradable polymers such as poly lactic acids, polyglycolic acids andany combination thereof, whether a physical blend or cross-linked. Incertain embodiments, the hydrogel components may be cross-linked. Thehydrogel forming excipient(s) are typically present at 0.1-70% w/w,preferably 1-50% w/w or most preferably 1-30% w/w.

In certain embodiments the formulations may also include at least onecontrolled release modifier which is a substance that upon hydration ofthe dosage form will preferentially adhere to the drug molecules andthus reduce the rate of its diffusion from the oral dosage form. Suchexcipients may also reduce the rate of water uptake by the formulationand thus enable a more prolonged drug dissolution and release from thetablet. In general the selected excipient(s) are lipophilic and capableof naturally complexing to the hydrophobic or lipophilic drugs. Thedegree of association of the release modifier and the drug can be variedby altering the modifier-to-drug ratio in the formulation. In addition,such interaction may be appropriately enhanced by the appropriatecombination of the release modifier with the active drug in themanufacturing process. Alternatively, the controlled release modifiermay be a charged polymer either synthetic or biopolymer bearing a netcharge, either positive or negative, and which is capable of binding tothe active via electrostatic interactions thus modifying both itsdiffusion through the tablet and/or the kinetics of its permeationthrough the mucosal surface. Similarly to the other compounds mentionedabove, such interaction is reversible and does not involve permanentchemical bonds with the active. In certain embodiments the controlledrelease modifier may typically be present at 0-80% w/w, preferably 1-20%w/w, most preferably 1-10% w/w.

In various embodiments the extended release formulations may alsoinclude other conventional components required for the development oforal dosage forms, which are known to those skilled in the art. Thesecomponents may include one or more bulking agents (such as lactose USP,Starch 1500, mannitol, sorbitol, malitol or other non-reducing sugars;microcrystalline cellulose (e.g., AVICEL®), dibasic calcium phosphatedehydrate, sucrose, and mixtures thereof), at least one solubilizingagent(s) (such as cyclodextrins, pH adjusters, salts and buffers,surfactants, fatty acids, phospholipids, metals of fatty acids etc.),metal salts and buffers organic (such as acetate, citrate, tartrate,etc.) or inorganic (phosphate, carbonate, bicarbonate, borate, sulfate,sulfite, bisulfite, metabisulfite, chloride, etc.), salts of metals suchas sodium, potassium, calcium, magnesium, etc.), at least one lubricant(such as stearic acid and divalent cations of, such as magnesiumstearate, calcium stearate, etc., talc, glycerol monostearate and thelike), one or more glidants (such as colloidal silicon dioxide,precipitated silicon dioxide, fumed silica (CAB-O-SIL® M-5P, trademarkof Cabot Corporation), stearowet and sterotex, silicas (such as SILOID®and SILOX® silicas—trademarks of Grace Davison Products,Aerosil—trademark of Degussa Pharma), higher fatty acids, the metalsalts thereof, hydrogenated vegetable oils and the like), flavors orsweeteners and colorants (such as aspartame, mannitol, lactose, sucrose,other artificial sweeteners; ferric oxides and FD&C lakes), additives tohelp stabilize the drug substance from chemical of physical degradation(such as anti-oxidants, anti-hydrolytic agents, aggregation-blockersetc. Anti-oxidants may include BHT, BHA, vitamins, citric acid, EDTA,sodium bisulfate, sodium metabisulfate, thiourea, methionine,surfactants, amino-acids, such as arginine, glycine, histidine,methionine salts, pH adjusters, chelating agents and buffers in the dryor solution form), one or more excipients that may affect tabletdisintegration kinetics and drug release from the tablet, and thuspharmacokinetics (disintegrants such as those known to those skilled inthe art and may be selected from a group consisting of starch,carboxy-methycellulose type or crosslinked polyvinyl pyrrolidone (suchas cross-povidone, PVP-XL), alginates, cellulose-based disintegrants(such as purified cellulose, methylcellulose, crosslinked sodium carboxymethylcellulose (Ac-Di-Sol) and carboxy methyl cellulose), lowsubstituted hydroxypropyl ethers of cellulose, microcrystallinecellulose (such as AVICEL®), ion exchange resins (such as AMBRELITE® IPR88), gums (such as agar, locust bean, karaya, pectin and tragacanth),guar gums, gum karaya, chitin and chitosan, smecta, gellan gum,isapghula husk, polacrillin potassium (Tulsion³³⁹)′ gas-evolvingdisintegrants (such as citric acid and tartaric acid along with thesodium bicarbonate, sodium carbonate, potassium bicarbonate or calciumcarbonate), sodium starch glycolate (such as EXPLOTAB® and PRIMOGEL®),starch DC and the likes, at least one biodegradable polymer of any typeuseful for extended drug release. Exemplary polymer compositionsinclude, but are not limited to, polyanhydrides and co-polymers oflactic acid and glycolic acid, poly(dl-lactide-co-glycolide) (PLGA),poly(lactic acid) (PLA), poly(glycolic acid) (PGA), polyorthoesters,proteins, and polysaccharides.

In certain embodiments, the active agent(s) can be chemically modifiedto significantly modify the pharmacokinetics in plasma. This may beaccomplished for example by conjugation with poly(ethylene glycol)(PEG), including site-specific PEGylation. PEGylation, which may improvedrug performance by optimizing pharmacokinetics, decreasingimmunogenicity and dosing frequency.

Methods of making a formulation of the active agent(s) described herein(e.g., triazolopyrimidine(s) and/or triazolopyridine(s) describedherein, or a tautomer(s) or stereoisomer(s) thereof, or pharmaceuticallyacceptable salts or solvates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) for GI or oral transmucosaldelivery are also provided. One method includes the steps of powdergrinding, dry powder mixing and tableting via direct compression.Alternatively, a wet granulation process may be used. Such a method(such as high shear granulation process) involves mixing the activeingredient and possibly some excipients in a mixer. The binder may beone of the excipients added in the dry mix state or dissolved in thefluid used for granulating. The granulating solution or suspension isadded to the dry powders in the mixer and mixed until the desiredcharacteristics are achieved. This usually produces a granule that willbe of suitable characteristics for producing dosage forms with adequatedissolution time, content uniformity, and other physicalcharacteristics. After the wet granulation step, the product is mostoften dried and/or then milled after drying to get a major percentage ofthe product within a desired size range. Sometimes, the product is driedafter being wet sized using a device such as an oscillating granulator,or a mill. The dry granulation may then processed to get an acceptablesize range by first screening with a sieving device, and then millingthe oversized particles.

Additionally, the formulation may be manufactured by alternativegranulation processes, all known to those skilled in the art, such asspray fluid bed granulation, extrusion and spheronization or fluid bedrotor granulation.

Additionally, the tablet dosage form of the active agent(s) describedherein may be prepared by coating the primary tablet manufactured asdescribed above with suitable coatings known in the art. Such coatingsare meant to protect the active cores against damage (abrasion,breakage, dust formation) against influences to which the cores areexposed during transport and storage (atmospheric humidity, temperaturefluctuations), and naturally these film coatings can also be colored.The sealing effect of film coats against water vapor is expressed by thewater vapor permeability. Coating may be performed by one of theavailable processes such as Wurster coating, dry coating, film coating,fluid bed coating, pan coating, etc. Typical coating materials includepolyvinyl pyrrolidone (PVP), polyvinyl pyrrolidone vinyl acetatecopolymer (PVPVA), polyvinyl alcohol (PVA), polyvinylalcohol/polyethylene glycol copolymer (PVA/PEG), cellulose acetatephthalate, ethyl cellulose, gellan gum, maltodextrin, methacrylates,methyl cellulose, hydroxyl propyl methyl cellulose (HPMC of all gradesand molecular weights), carrageenan, shellac and the like.

In certain embodiments the tablet core comprising the active agent(s)described herein can be coated with a bioadhesive and/or pH resistantmaterial to enable material, such as those defined above, to improvebioadhesion of the tablet in the sublingual cavity.

In certain embodiments, the active agent(s) described herein (e.g.,triazolopyrimidine(s) and/or triazolopyridine(s) described herein, or atautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts or solvates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) are formulated as inclusioncomplexes. While not limited to cyclodextrin inclusion complexes, it isnoted that cyclodextrin is the agent most frequently used to formpharmaceutical inclusion complexes. Cyclodextrins (CD) are cyclicoligomers of glucose, that typically contain 6, 7, or 8 glucose monomersjoined by α-1,4 linkages. These oligomers are commonly called α-CD,β-CD, and γ-CD, respectively. Higher oligomers containing up to 12glucose monomers are known, and contemplated to in the formulationsdescribed herein. Functionalized cyclodextrin inclusion complexes arealso contemplated. Illustrative, but non-limiting functionalizedcyclodextrins include, but are not limited to sulfonates, sulfonates andsulfinates, or disulfonates of hydroxybutenyl cyclodextrin; sulfonates,sulfonates and sulfinates, or disulfonates of mixed ethers ofcyclodextrins where at least one of the ether substituents ishydroxybutenyl cyclodextrin. Illustrative cyclodextrins include apolysaccharide ether which comprises at least one 2-hydroxybutenylsubstituent, wherein the at least one hydroxybutenyl substituent issulfonated and sulfinated, or disulfonated, and an alkylpolyglycosideether which comprises at least one 2-hydroxybutenyl substituent, whereinthe at least one hydroxybutenyl substituent is sulfonated andsulfinated, or disulfonated. In various embodiments inclusion complexesformed between sulfonated hydroxybutenyl cyclodextrins and one or moreof the active agent(s) described herein are contemplated. Methods ofpreparing cyclodextrins, and cyclodextrin inclusion complexes are foundfor example in U.S. Patent Publication No: 2004/0054164 and thereferences cited therein and in U.S. Patent Publication No: 2011/0218173and the references cited therein.

Pharmacokinetics (PK) and Formulation Attributes

One advantage of the extended (controlled) release oral (GI ortransmucosal) formulations described herein is that they can maintainthe plasma drug concentration within a targeted therapeutic window for alonger duration than with immediate-release formulations, whether soliddosage forms or liquid-based dosage forms. The high peak plasma levelstypically observed for such conventional immediate release formulationswill be blunted by the prolonged release of the drug over 1 to 12 hoursor longer. In addition, a rapid decline in plasma levels will be avoidedsince the drug will continually be crossing from the oral cavity intothe bloodstream during the length of time of dissolution of the tablet,thus providing plasma pharmacokinetics with a more stable plateau. Inaddition, the dosage forms described herein may improve treatment safetyby minimizing the potentially deleterious side effects due to thereduction of the peaks and troughs in the plasma drug pharmacokinetics,which compromise treatment safety.

In various embodiments the oral transmucosal formulations of the activeagent(s) described herein designed to avoid the high peak plasma levelsof intravenous and conventional immediate release oral dosage forms byutilizing the mucosa and by independently controlling both tabletdisintegration (or erosion) and drug dissolution and release from thetablet over time to provide a safer delivery profile. The oralformulations described herein provide individual, repetitive doses thatinclude a defined amount of the active agent.

An advantage of the bioadhesive oral transmucosal formulations describedherein is that they exhibit highly consistent bioavailability and canmaintain the plasma drug concentration within a targeted therapeuticwindow with significantly lower variability for a longer duration thancurrently available dosage forms, whether solid dosage forms or IVdosage forms. In addition, a rapid decline in plasma levels is avoidedsince the drug is continually crossing from the oral cavity or GI tractinto the bloodstream during the length of time of dissolution of thetablet or longer, thus providing plasma pharmacokinetics with anextended plateau phase as compared to the conventional immediate releaseoral dosage forms. Further, the dosage forms described herein canimprove treatment safety by minimizing the potentially deleterious sideeffects due to the relative reduction of the peaks and troughs in theplasma drug pharmacokinetics, which compromise treatment safety and istypical of currently available dosage forms.

In various embodiments bioadhesive formulations described herein can bedesigned to manipulate and control the pharmacokinetic profile of theactive agent(s) described herein. As such, the formulations can beadjusted to achieve ‘slow’ disintegration times (and erosion kineticprofiles) and slow drug release and thus enable very prolongedpharmacokinetic profiles that provide sustained drug action. Althoughsuch formulations may be designed to still provide a fast onset, theyare mostly intended to enable the sustained drug PK and effect whilemaintaining the other performance attributes of the tablet such asbioadhesion, reproducibility of action, blunted C_(max), etc.

The performance and attributes of the bioadhesive transmucosalformulations of this invention are independent of the manufacturingprocess. A number of conventional, well-established and known in the artprocesses can be used to manufacture the formulations of the presentinvention (such as wet and dry granulation, direct compression, etc.)without impacting the dosage form physicochemical properties or in vivoperformance.

An illustrative mathematical ratio that demonstrates the prolongedplateau phase of the measured blood plasma levels of the active agent(s)described herein, following administration of the dosage forms of theinvention is the term “Optimal Therapeutic Targeting Ratio” or “OTTR”,which represents the average time that the drug is present attherapeutic levels, defined as time within which the drug plasmaconcentration is maintained above 50% of C_(max) normalized by thedrug's elimination half-life multiplied by the ratio of the C_(max)obtained in the dosage form of interest over the normalized C_(max)following IV administration of equivalent doses. In certain embodimentsthe OTTR can be calculated by the formula:

OTTR=(C ^(IV) _(max) /C _(max))×(Dose/Dose^(IV))(Time above 50% of C_(max))/(Terminal^(IV) elimination half-life of the drug).

In certain embodiments the OTTR is greater than about 15, or greaterthan about 20, or greater than about 25, or greater than about 30, orgreater than about 40, or greater than about 50.

Administration

In certain embodiments one or more active agents described herein (e.g.,triazolopyrimidine(s) and/or triazolopyridine(s) described herein, or atautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts or solvates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) are administered to amammal in need thereof, e.g., to a mammal at risk for or suffering froma pathology characterized by abnormal processing of amyloid precursorproteins, a mammal at risk for progression of MCI to Alzheimer'sdisease, and so forth. In certain embodiments the active agent(s) areadministered to prevent or delay the onset of a pre-Alzheimer'scognitive dysfunction, and/or to ameliorate one or more symptoms of apre-Alzheimer's cognitive dysfunction, and/or to prevent or delay theprogression of a pre-Alzheimer's condition or cognitive dysfunction toAlzheimer's disease, and/or to promote the processing of amyloidprecursor protein (APP) by a non-amyloidogenic pathway. In certainembodiments one or more active agent(s) are administered for thetreatment of early stage, mid stage, or late-stage Alzheimer's disease,e.g., to reduce the severity of the disease, and/or to ameliorate one ormore symptoms of the disease, and/or to slow the progression of thedisease.

In various embodiments the active agent(s) described herein (e.g.,triazolopyrimidine(s) and/or triazolopyridine(s) described herein, or atautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts or solvates of said triazolopyrimidine(s) and/ortriazolopyridine(s), said stereoisomer(s), or said tautomer(s), oranalogues, derivatives, or prodrugs thereof) can be administered by anyof a number of routes. Thus, for example they can be administeredorally, parenterally, (intravenously (IV), intramuscularly (IM),depo-IM, subcutaneously (SQ), and depo-SQ), sublingually, intranasally(inhalation), intrathecally, transdermally (e.g., via transdermalpatch), topically, ionophoretically or rectally. Typically the dosageform is selected to facilitate delivery to the brain (e.g., passagethrough the blood brain barrier). In this context it is noted that thecompounds described herein are readily delivered to the brain. Dosageforms known to those of skill in the art are suitable for delivery ofthe compound.

In various embodiments the active agent(s) are administered in anamount/dosage regimen sufficient to exert a prophylactically and/ortherapeutically useful effect in the absence of undesirable side effectson the subject treated (or with the presence of acceptable levels and/ortypes of side effects). The specific amount/dosage regimen will varydepending on the weight, gender, age and health of the individual; theformulation, the biochemical nature, bioactivity, bioavailability andthe side effects of the particular compound.

In certain embodiments the therapeutically or prophylactically effectiveamount may be determined empirically by testing the agent(s) in known invitro and in vivo model systems for the treated disorder. Atherapeutically or prophylactically effective dose can be determined byfirst administering a low dose, and then incrementally increasing untila dose is reached that achieves the desired effect with minimal or noundesired side effects.

In certain embodiments, when administered orally, an administered amountof the agent(s) described herein effective to prevent or delay the onsetof a pre-Alzheimer's cognitive dysfunction, and/or to ameliorate one ormore symptoms of a pre-Alzheimer's cognitive dysfunction, and/or toprevent or delay the progression of a pre-Alzheimer's condition orcognitive dysfunction to Alzheimer's disease, and/or to promote theprocessing of amyloid precursor protein (APP) by a non-amyloidogenicpathway, and/or to treat or prevent AD ranges from about 0.1 mg/day toabout 500 mg/day or about 1,000 mg/day, or from about 0.1 mg/day toabout 200 mg/day, for example, from about 1 mg/day to about 100 mg/day,for example, from about 5 mg/day to about 50 mg/day. In someembodiments, the subject is administered the compound at a dose of about0.05 to about 0.50 mg/kg, for example, about 0.05 mg/kg, 0.10 mg/kg,0.20 mg/kg, 0.33 mg/kg, 0.50 mg/kg. It is understood that while apatient may be started at one dose, that dose may be varied (increasedor decreased, as appropriate) over time as the patient's conditionchanges. Depending on outcome evaluations, higher doses may be used. Forexample, in certain embodiments, up to as much as 1000 mg/day can beadministered, e.g., 5 mg/day, 10 mg/day, 25 mg/day, 50 mg/day, 100mg/day, 200 mg/day, 300 mg/day, 400 mg/day, 500 mg/day, 600 mg/day, 700mg/day, 800 mg/day, 900 mg/day or 1000 mg/day.

In various embodiments, active agent(s) described herein can beadministered parenterally, for example, by IV, IM, depo-IM, SC, ordepo-SC. In certain embodiments when administered parenterally, atherapeutically effective amount of about 0.5 to about 100 mg/day,preferably from about 5 to about 50 mg daily can be delivered. When adepot formulation is used for injection once a month or once every twoweeks, the dose in certain embodiments can be about 0.5 mg/day to about50 mg/day, or a monthly dose of from about 15 mg to about 1,500 mg. Inpart because of the forgetfulness of the patients with Alzheimer'sdisease, it is preferred that the parenteral dosage form be a depoformulation.

In various embodiments, the active agent(s) described herein can beadministered sublingually. In some embodiments, when given sublingually,the compounds and/or analogs thereof can be given one to four timesdaily in the amounts described above for IM administration.

In various embodiments, the active agent(s) described herein can beadministered intranasally. When given by this route, the appropriatedosage forms are a nasal spray or dry powder, as is known to thoseskilled in the art. In certain embodiments, the dosage of compoundand/or analog thereof for intranasal administration is the amountdescribed above for IM administration.

In various embodiments, the active agent(s) described herein can beadministered intrathecally. When given by this route the appropriatedosage form can be a parenteral dosage form as is known to those skilledin the art. In certain embodiments, the dosage of compound and/or analogthereof for intrathecal administration is the amount described above forIM administration.

In certain embodiments, the active agent(s) described herein can beadministered topically. When given by this route, the appropriate dosageform is a cream, ointment, or patch. When administered topically, thedosage is from about 1.0 mg/day to about 200 mg/day. Because the amountthat can be delivered by a patch is limited, two or more patches may beused. The number and size of the patch is not important as long as atherapeutically effective amount of compound be delivered as is known tothose skilled in the art. The compound can be administered rectally bysuppository as is known to those skilled in the art. In certainembodiments, when administered by suppository, the therapeuticallyeffective amount is from about 1.0 mg to about 500 mg.

In various embodiments, the active agent(s) described herein can beadministered by implants as is known to those skilled in the art. Whenadministering the compound by implant, the therapeutically effectiveamount is the amount described above for depot administration.

In various embodiments, the active agent(s) described herein thereof canbe enclosed in multiple or single dose containers. The enclosed agent(s)can be provided in kits, for example, including component parts that canbe assembled for use. For example, an active agent in lyophilized formand a suitable diluent may be provided as separated components forcombination prior to use. A kit may include an active agent and a secondtherapeutic agent for co-administration. The active agent and secondtherapeutic agent may be provided as separate component parts. A kit mayinclude a plurality of containers, each container holding one or moreunit dose of the compounds. The containers are preferably adapted forthe desired mode of administration, including, but not limited totablets, gel capsules, sustained-release capsules, and the like for oraladministration; depot products, pre-filled syringes, ampules, vials, andthe like for parenteral administration; and patches, medipads, creams,and the like for topical administration, e.g., as described herein.

In various embodiments the dosage forms can be administered to thesubject 1, 2, 3, or 4 times daily. In certain embodiments it ispreferred that the compound be administered either three or fewer times,more preferably once or twice daily. In certain embodiments, it ispreferred that the agent(s) be administered in oral dosage form.

It should be apparent to one skilled in the art that the exact dosageand frequency of administration will depend on the particular conditionbeing treated, the severity of the condition being treated, the age,weight, general physical condition of the particular patient, and othermedication the individual may be taking as is well known toadministering physicians who are skilled in this art.

While the compositions and methods are described herein with respect touse in humans, they are also suitable for animal, e.g., veterinary use.Thus certain organisms (subjects) contemplated herein include, but arenot limited to humans, non-human primates, canines, equines, felines,porcines, ungulates, largomorphs, and the like.

The foregoing formulations and administration methods are intended to beillustrative and not limiting. It will be appreciated that, using theteaching provided herein, other suitable formulations and modes ofadministration can be readily devised.

Combination Therapies

In certain embodiments, the active agent(s) described herein (e.g.,triazolopyrimidine(s) and/or triazolopyridine(s) described herein, or atautomer(s) or stereoisomer(s) thereof, or pharmaceutically acceptablesalts, solvates, or clathrates of said compounds, said stereoisomer(s),or said tautomer(s), or analogues, derivatives, or prodrugs thereof) canbe used in combination with other therapeutic agents or approaches usedto treat or prevent diseases characterized by amyloid deposits in thebrain, including MCI and/or AD. Accordingly, in certain embodiments, apharmaceutical composition comprising at least one active agentdescribed herein (e.g., a triazolopyrimidine and/or triazolopyridinedescribed herein, or a tautomer or stereoisomer thereof, orpharmaceutically acceptable salt, solvate, or clathrate of saidtriazolopyrimidine and/or triazolopyridine, said stereoisomer, or saidtautomer, or an analogue, derivative, or prodrug thereof) one togetherwith at least one additional therapeutic agent, and, optionally, apharmaceutically acceptable carrier or diluent is contemplated. Incertain embodiments a therapeutic or prophylactic method comprisingadministering at least active agent described herein in conjunction withat least one additional therapeutic agent is contemplated.

In certain embodiments non-limiting examples of additional therapeuticagents include, but are not limited to disulfiram and/or analoguesthereof, honokiol and/or analogues thereof, tropisetron and/or analoguesthereof, nimetazepam and/or analogues thereof (see, e.g., U.S. Ser. No.13/213,960 (U.S. Patent Publication No: US-2012-0071468-A1), andPCT/US2011/048472 (PCT Publication No: WO 2012/024616) which areincorporated herein by reference for the compounds described therein),tropinol-esters and/or related esters and/or analogues thereof (see,e.g., U.S. Ser. 61/514,381, which is incorporated herein by referencefor the compounds described herein), TrkA kinase inhibitors (e.g.,ADDN-1351) and/or analogues thereof (see, e.g., U.S. Ser. No.61/525,076, which is incorporated herein by reference for the compoundsdescribed therein), hydantoins and/or analogues thereof (see, e.g.,PCT/US2014/016100 (WO 2014/127042 A1), which is incorporated herein byreference for the compounds described therein, D2 receptor agonists andalpha1-adrenergic receptor antagonists, and APP-specific BACE Inhibitors(ASBIs) as described and/or claimed in PCT/US2013/032481 (WO 2013/142370A1) and U.S. Ser. No. 14/384,641 which are incorporated herein byreference for the active agents described therein including, but notlimited to galangin, a galangin prodrug, rutin, a rutin prodrug, andother flavonoids and flavonoid prodrugs as described or claimed therein.

Non-limiting examples of additional therapeutic agents include drugsselected from the group consisting of: (a) drugs useful for thetreatment of Alzheimer's disease and/or drugs useful for treating one ormore symptoms of Alzheimer's disease, (b) drugs useful for inhibitingthe synthesis Aβ, and (c) drugs useful for treating neurodegenerativediseases. Additional non-limiting examples of additional therapeuticagents for use in combination with the compounds (e.g.,triazolopyrimidine and/or triazolopyridine) described herein includedrugs useful for the treatment, prevention, delay of onset, ameliorationof any pathology associated with Aβ and/or a symptom thereof.Non-limiting examples of pathologies associated with Aβ include:Alzheimer's disease, Down's syndrome, Parkinson's disease, memory loss,memory loss associated with Alzheimer's disease, memory loss associatedwith Parkinson's disease, attention deficit symptoms, attention deficitsymptoms associated with Alzheimer's disease, Parkinson's disease,and/or Down's syndrome, dementia, stroke, microgliosis and braininflammation, pre-senile dementia, senile dementia, dementia associatedwith Alzheimer's disease, Parkinson's disease, and/or Down's syndrome,progressive supranuclear palsy, cortical basal degeneration,neurodegeneration, olfactory impairment, olfactory impairment associatedwith Alzheimer's disease, Parkinson's disease, and/or Down's syndrome,β-amyloid angiopathy, cerebral amyloid angiopathy, hereditary cerebralhemorrhage, mild cognitive impairment (“MCI”), glaucoma, amyloidosis,type II diabetes, hemodialysis complications (from (3.sub.2microglobulins and complications arising therefrom in hemodialysispatients), scrapie, bovine spongiform encephalitis, traumatic braininjury (“TBI”), and Creutzfeld-Jakob disease, comprising administeringto said patient at least one triazolopyrimidine and/or triazolopyridinecompound described herein, or a tautomer or isomer thereof orpharmaceutically acceptable salt or solvate of said compound or saidtautomer, in an amount effective to inhibit said pathology orpathologies.

In certain embodiments such additional therapeutic agents include, butare not limited to acetylcholinesterase inhibitors (including withoutlimitation, e.g., (−)-phenserine enantiomer, tacrine, ipidacrine,galantamine, donepezil, icopezil, zanapezil, rivastigmine, huperzine A,phenserine, physostigmine, neostigmine, pyridostigmine, ambenonium,demarcarium, edrophonium, ladostigil and ungeremine); NMDA receptorantagonist (including without limitations e.g., Memantine); muscarinicreceptor agonists (including without limitation, e.g., Talsaclidine,AF-102B, AF-267B (NGX-267)); nicotinic receptor agonists (includingwithout limitation, e.g., Ispronicline (AZD-3480)); beta-secretaseinhibitors (including without limitations e.g., thiazolidinediones,including rosiglitazone and pioglitazone); gamma-secretase inhibitors(including without limitation, e.g., semagacestat (LY-450139), MK-0752,E-2012, BMS-708163, PF-3084014, begacestat (GSI-953), and NIC5-15);inhibitors of Aβ aggregation (including without limitation, e.g.,Clioquinol (PBT1), PBT2, tramiprosate (homotaurine), Scyllo-inositol(a.k.a., scyllo-cyclohexanehexol, AZD-103 and ELND-005), passiveimmunotherapy with Aβ fragments (including without limitations e.g.,Bapineuzemab) and Epigallocatechin-3-gallate (EGCg)); anti-inflammatoryagents such as cyclooxygenase II inhibitors; anti-oxidants such asVitamin E and ginkolides; immunological approaches, such as, forexample, immunization with Aβ peptide or administration of anti-Aβpeptide antibodies; statins; and direct or indirect neurotrophic agentssuch as Cerebrolysin™, AIT-082 (Emilieu, 2000, Arch. Neurol. 57:454),Netrin (Luorenco (2009) Cell Death Differ., 16: 655-663), Netrinmimetics, NGF, NGF mimetics, BDNF and other neurotrophic agents of thefuture, agents that promote neurogenesis e.g. stem cell therapy. Furtherpharmacologic agents useful in the treatment or prevention diseasescharacterized by amyloid deposits in the brain, including MCI and/or AD,are described, e.g., in Mangialasche et al. (2010) Lancet Neurol., 9:702-716.

In certain embodiments, additional non-limiting examples of additionaltherapeutic agents for use in combination with compounds describedherein include: muscarinic antagonists (e.g., m₁ agonists (such asacetylcholine, oxotremorine, carbachol, or McNa343), or m₂ antagonistscholinesterase inhibitors (e.g., acetyl- and/or butyrylchlolinesteraseinhibitors such as donepezil (Aricept®), galantamine (RAZADYNE®), andrivastigimine (EXELON®); N-methyl-D-aspartate receptor antagonists(e.g., NAMENDA® (memantine HCl); combinations of cholinesteraseinhibitors and N-methyl-D-aspartate receptor antagonists; gammasecretase modulators; gamma secretase inhibitors; non-steroidalanti-inflammatory agents; anti-inflammatory agents that can reduceneuroinflammation; anti-amyloid antibodies (such as bapineuzemab,Wyeth/Elan); vitamin E; nicotinic acetylcholine receptor agonists; CB1receptor inverse agonists or CB1 receptor antagonists; antibiotics;growth hormone secretagogues; histamine H3 antagonists; AMPA agonists;PDE4 inhibitors; GABA_(A) inverse agonists; inhibitors of amyloidaggregation; glycogen synthase kinase beta inhibitors; promoters ofalpha secretase activity; PDE-10 inhibitors; Tau kinase inhibitors(e.g., GSK3beta inhibitors, cdk5 inhibitors, or ERK inhibitors); Tauaggregation inhibitors (e.g., REMBER®; RAGE inhibitors (e.g., TTP 488(PF-4494700)); anti-Aβ vaccine; APP ligands; agents that upregulateinsulin, cholesterol lowering agents such as HMG-CoA reductaseinhibitors (for example, statins such as Atorvastatin, Fluvastatin,Lovastatin, Mevastatin, Pitavastatin, Pravastatin, Rosuvastatin,Simvastatin) and/or cholesterol absorption inhibitors (such asEzetimibe), or combinations of HMG-CoA reductase inhibitors andcholesterol absorption inhibitors (such as, for example, VYTORIN®);fibrates (such as, for example, clofibrate, Clofibride, Etofibrate, andAluminium Clofibrate); combinations of fibrates and cholesterol loweringagents and/or cholesterol absorption inhibitors; nicotinic receptoragonists; niacin; combinations of niacin and cholesterol absorptioninhibitors and/or cholesterol lowering agents (e.g., SIMCOR®(niacin/simvastatin, available from Abbott Laboratories, Inc.); LXRagonists; LRP mimics; H3 receptor antagonists; histone deacetylaseinhibitors; hsp90 inhibitors; 5-HT4 agonists (e.g., PRX-03140 (EpixPharmaceuticals)); 5-HT6 receptor antagonists; mGluR1 receptormodulators or antagonists; mGluR5 receptor modulators or antagonists;mGluR2/3 antagonists; Prostaglandin EP2 receptor antagonists; PAI-1inhibitors; agents that can induce Abeta efflux such as gelsolin;Metal-protein attenuating compound (e.g., PBT2); and GPR3 modulators;and antihistamines such as Dimebolin (e.g., DIMEBON®, Pfizer).

Accordingly certain embodiments provide a pharmaceutical compositioncomprising an effective amount of one or more triazolopyrimidine(s)and/or triazolopyridine(s) described herein and an additionaltherapeutic agent, and/or a method of treatment or prophylaxiscomprising administration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is disulfiram and/or analogues thereof (see, e.g., U.S. Ser. No.13/213,960 (U.S. Patent Publication No: US-2012-0071468-A1), andPCT/US2011/048472 (PCT Publication No: WO 2012/024616) which areincorporated herein by references for the compounds described therein).

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is honokiol and/or analogues thereof (see, e.g., U.S. Ser. No.13/213,960 (U.S. Patent Publication No: US-2012-0071468-A1), andPCT/US2011/048472 (PCT Publication No: WO 2012/024616) which areincorporated herein by references for the compounds described therein).

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is tropisetron and/or analogues thereof (see, e.g., U.S. Ser. No.13/213,960 (U.S. Patent Publication No: US-2012-0071468-A1), andPCT/US2011/048472 (PCT Publication No: WO 2012/024616) which areincorporated herein by references for the compounds described therein).

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is tropisetron.

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is nimetazepam and/or analogues thereof (see, e.g., U.S. Ser. No.13/213,960 (U.S. Patent Publication No: US-2012-0071468-A1), andPCT/US2011/048472 (PCT Publication No: WO 2012/024616) which areincorporated herein by references for the compounds described therein).

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is a tropinol ester or related ester (see, e.g.,PCT/US2012/049223 (WO 2013/019901 A2), and U.S. Ser. No. 14/235,405which are incorporated herein by reference for the tropinol esters andrelated compounds described therein).

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is a TrkA kinase inhibitor (e.g., ADDN-1351) and/or analoguesthereof (see, e.g., PCT/US2012/051426 (WO 2013/026021 A2), which isincorporated hereby reference for the TrkA kinase inhibitors and analogsthereof described therein.

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is a D2 receptor agonists and/or an alpha1-adrenergic receptorantagonists.

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is an ASBIs as described and/or claimed in PCT/US2013/032481 (WO2013/142370 A1) and U.S. Ser. No. 14/384,641 which are incorporatedherein by reference for the active agents described therein including,but not limited to galangin, a galangin prodrug, rutin, and otherflavonoids as described or claimed therein.

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is one or more cholinesterase inhibitors (e.g., acetyl- and/orbutyrylchlolinesterase inhibitors).

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is one or more muscarinic antagonists (e.g., m₁ agonists or m₂antagonists).

Certain embodiments provide a pharmaceutical composition comprising aneffective amount of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein and an additional therapeuticagent, and/or a method of treatment or prophylaxis comprisingadministration of one or more triazolopyrimidine(s) and/ortriazolopyridine(s) described herein in conjunction with an additionaltherapeutic agent where the therapeutic agent in the formulation and/ormethod is one or more compounds selected from the group consisting ofcholinesterase inhibitors (such as, for example,(.+−.)-2,3-dihydro-5,6-dimethoxy-2-[[1-(phenylmethyl)-4-piperidinyl]methy-1]-1H-inden-1-onehydrochloride, i.e, donepezil hydrochloride, available as the ARICEPT®brand of donepezil hydrochloride), N-methyl-D-aspartate receptorinhibitors (such as, for example, Namenda® (memantine HCl));anti-amyloid antibodies (such as bapineuzumab, Wyeth/Elan), gammasecretase inhibitors, gamma secretase modulators, and beta secretaseinhibitors other than the triazolopyrimidine(s) and/ortriazolopyridine(s) described herein.

Methods of Monitoring Clinical Efficacy

In various embodiments, the effectiveness of treatment can be determinedby comparing a baseline measure of a parameter of disease beforeadministration of the agent(s) (e.g., triazolopyrimidine(s) and/ortriazolopyridine(s) described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts orsolvates of said triazolopyrimidine and/or triazolopyridine(s), saidstereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof) is commenced to the same parameter one or more timepoints after the agent(s) or analog has been administered. Oneillustrative parameter that can be measured is a biomarker (e.g., apeptide oligomer) of APP processing or related and implicated pathways.Such biomarkers include, but are not limited to sAPPα, p3 (A1317-42 orAβ17-40), sAPPβ, soluble Aβ40, and/or soluble Aβ42 in the blood, plasma,serum, urine, mucous or cerebrospinal fluid (CSF).

An important indicator is p-tau and tau. A reduction of p-tau and/or tauis an indication of desirable efficacy. Additionally, inhibition inelevation of p-tau, e.g., stress-induced (cortisol-induced) p-tau isalso an indication of efficacy.

Additionally, detection of increased levels of sAPPα and/or p3, anddecreased levels of sAPPβ and/or APPneo is an indicator that thetreatment is effective. Conversely, detection of decreased levels ofsAPPα and/or p3, and/or increased levels of sAPPβ, APPneo, Tau orphospho-Tau (pTau) may be an indicator that the treatment is noteffective.

Another parameter to determine effectiveness of treatment is the levelof amyloid plaque deposits in the brain. Amyloid plaques can bedetermined using any method known in the art, e.g., as determined by CT,PET, PIB-PET and/or MRI. Administration of one or more of the agent(s))described herein (e.g., triazolopyrimidine(s) and/ortriazolopyridine(s), or a tautomer(s) or stereoisomer(s) thereof, orpharmaceutically acceptable salt, solvate, or clathrate saidcompound(s), said stereoisomer(s), or said tautomer(s), or analogues,derivatives, or prodrugs thereof) can result in a reduction in the rateof plaque formation, and even a retraction or reduction of plaquedeposits in the brain. Effectiveness of treatment can also be determinedby observing a stabilization and/or improvement of cognitive abilitiesof the subject. Cognitive abilities can be evaluated using anyart-accepted method, including for example, Clinical Dementia Rating(CDR), the mini-mental state examination (MMSE) or Folstein test,evaluative criteria listed in the DSM-IV (Diagnostic and StatisticalManual of Mental Disorders, Fourth Edition) or DSM-V, and the like.

Clinical efficacy can be monitored using any method known in the art.Measurable biomarkers to monitor efficacy include, but are not limitedto, monitoring blood, plasma, serum, urine, mucous or cerebrospinalfluid (CSF) levels of sAPPα, sAPPβ, Aβ42, Aβ40, APPneo and p3 (e.g.,Aβ17-42 or Aβ17-40). Detection of increased levels of sAPPα and/or p3,and decreased levels of sAPPβ and/or APPneo are indicators that thetreatment or prevention regime is efficacious. Conversely, detection ofdecreased levels of sAPPα and/or p3, and increased levels of sAPPβand/or APPneo are indicators that the treatment or prevention regime isnot efficacious. Other biomarkers include Tau and phospho-Tau (pTau).Detection of decreased levels of Tau and pTau are indicators that thetreatment or prevention regime is efficacious.

Efficacy can also be determined by measuring amyloid plaque load in thebrain. The treatment or prevention regime is considered efficacious whenthe amyloid plaque load in the brain does not increase or is reduced.Conversely, the treatment or prevention regime is consideredinefficacious when the amyloid plaque load in the brain increases.Amyloid plaque load can be determined using any method known in the art,e.g., including CT, PET, PIB-PET and/or MM.

Efficacy can also be determined by measuring the cognitive abilities ofthe subject. Cognitive abilities can be measured using any method knownin the art. Illustrative tests include assigning a Clinical DementiaRating (CDR) score or applying the mini mental state examination (MMSE)(Folstein, et al. (1975) J Psychiatric Res. 12(3): 189-198). Subjectswho maintain the same score or who achieve an improved score, e.g., whenapplying the CDR or MMSE, indicate that the treatment or preventionregime is efficacious. Conversely, subjects who receive a scoreindicating diminished cognitive abilities, e.g., when applying the CDRor MMSE, indicate that the treatment or prevention regime has not beenefficacious.

In certain embodiments, the monitoring methods can entail determining abaseline value of a measurable biomarker or parameter (e.g., amyloidplaque load or cognitive abilities) in a subject before administering adosage of the agent(s), and comparing this with a value for the samemeasurable biomarker or parameter after treatment.

In other methods, a control value (e.g., a mean and standard deviation)of the measurable biomarker or parameter is determined for a controlpopulation. In certain embodiments, the individuals in the controlpopulation have not received prior treatment and do not have AD, MCI,nor are at risk of developing AD or MCI. In such cases, if the value ofthe measurable biomarker or clinical parameter approaches the controlvalue, then treatment is considered efficacious. In other embodiments,the individuals in the control population have not received priortreatment and have been diagnosed with AD or MCI. In such cases, if thevalue of the measurable biomarker or clinical parameter approaches thecontrol value, then treatment is considered inefficacious.

In other methods, a subject who is not presently receiving treatment buthas undergone a previous course of treatment is monitored for one ormore of the biomarkers or clinical parameters to determine whether aresumption of treatment is required. The measured value of one or moreof the biomarkers or clinical parameters in the subject can be comparedwith a value previously achieved in the subject after a previous courseof treatment. Alternatively, the value measured in the subject can becompared with a control value (mean plus standard deviation/ANOVA)determined in population of subjects after undergoing a course oftreatment. Alternatively, the measured value in the subject can becompared with a control value in populations of prophylactically treatedsubjects who remain free of symptoms of disease, or populations oftherapeutically treated subjects who show amelioration of diseasecharacteristics. In such cases, if the value of the measurable biomarkeror clinical parameter approaches the control value, then treatment isconsidered efficacious and need not be resumed. In all of these cases, asignificant difference relative to the control level (e.g., more than astandard deviation) is an indicator that treatment should be resumed inthe subject.

In certain embodiments the tissue sample for analysis is typicallyblood, plasma, serum, urine, mucous or cerebrospinal fluid from thesubject.

Kits.

In various embodiments, the active agent(s) (e.g., triazolopyrimidine(s)and/or triazolopyridine(s)) described herein thereof can be enclosed inmultiple or single dose containers. The enclosed agent(s) can beprovided in kits, for example, including component parts that can beassembled for use. For example, an active agent in lyophilized form anda suitable diluent may be provided as separated components forcombination prior to use. A kit may include an active agent and a secondtherapeutic agent for co-administration. The active agent and secondtherapeutic agent may be provided as separate component parts. A kit mayinclude a plurality of containers, each container holding one or moreunit dose of the compounds. The containers are preferably adapted forthe desired mode of administration, including, but not limited totablets, gel capsules, sustained-release capsules, and the like for oraladministration; depot products, pre-filled syringes, ampules, vials, andthe like for parenteral administration; and patches, medipads, creams,and the like for topical administration, e.g., as described herein.

In certain embodiments, a kit is provided where the kit comprises one ormore triazolopyrimidine and/or triazolopyridine compounds describedherein, or a tautomer or stereoisomer thereof, or pharmaceuticallyacceptable salt, solvate, or clathrate of said compound, saidstereoisomer, or said tautomer, preferably provided as a pharmaceuticalcomposition and in a suitable container or containers and/or withsuitable packaging; optionally one or more additional active agents,which if present are preferably provided as a pharmaceutical compositionand in a suitable container or containers and/or with suitablepackaging; and optionally instructions for use, for example writteninstructions on how to administer the compound or compositions.

In another embodiment, a kit is provided that comprises a singlecontainer or multiple containers: (a) a pharmaceutically acceptablecomposition comprising one or more compounds described herein (e.g.,compounds shown in FIG. 2, or illustrated or described in Table 5, or atautomer or stereoisomer thereof, or pharmaceutically acceptable salt,solvate, or clathrate of said compound, said stereoisomer, or saidtautomer, optionally a pharmaceutically acceptable compositioncomprising one or more additional therapeutic agents; and optionallyinstructions for use their use. The kit may optionally comprise labeling(e.g., instructional materials) appropriate to the intended use or uses.

As with any pharmaceutical product, the packaging material(s) and/orcontainer(s) are designed to protect the stability of the product duringstorage and shipment. In addition, the kits can include instructions foruse or other informational material that can advise the user such as,for example, a physician, technician or patient, regarding how toproperly administer the composition(s) as prophylactic, therapeutic, orameliorative treatment of the disease of concern. In some embodiments,instructions can indicate or suggest a dosing regimen that includes, butis not limited to, actual doses and monitoring procedures.

In some embodiments, the instructions can include informational materialindicating that the administering of the compositions can result inadverse reactions including but not limited to allergic reactions suchas, for example, anaphylaxis. The informational material can indicatethat allergic reactions may exhibit only as mild pruritic rashes or maybe severe and include erythroderma, vasculitis, anaphylaxis,Steven-Johnson syndrome, and the like. In certain embodiments theinformational material(s) may indicate that anaphylaxis can be fatal andmay occur when any foreign protein is introduced into the body. Incertain embodiments the informational material may indicate that theseallergic reactions can manifest themselves as urticaria or a rash anddevelop into lethal systemic reactions and can occur soon after exposuresuch as, for example, within 10 minutes. The informational material canfurther indicate that an allergic reaction may cause a subject toexperience paresthesia, hypotension, laryngeal edema, mental statuschanges, facial or pharyngeal angioedema, airway obstruction,bronchospasm, urticaria and pruritus, serum sickness, arthritis,allergic nephritis, glomerulonephritis, temporal arthritis,eosinophilia, or a combination thereof.

While the instructional materials typically comprise written or printedmaterials they are not limited to such. Any medium capable of storingsuch instructions and communicating them to an end user is contemplatedherein. Such media include, but are not limited to electronic storagemedia (e.g., magnetic discs, tapes, cartridges, chips), optical media(e.g., CD ROM), and the like. Such media may include addresses tointernet sites that provide such instructional materials.

In some embodiments, the kits can comprise one or more packagingmaterials such as, for example, a box, bottle, tube, vial, container,sprayer, insufflator, intravenous (I.V.) bag, envelope, and the like;and at least one unit dosage form of an agent comprising active agent(s)described herein and a packaging material. In some embodiments, the kitsalso include instructions for using the composition as prophylactic,therapeutic, or ameliorative treatment for the disease of concern.

In some embodiments, the articles of manufacture can comprise one ormore packaging materials such as, for example, a box, bottle, tube,vial, container, sprayer, insufflator, intravenous (IV.) bag, envelope,and the like; and a first composition comprising at least one unitdosage form of an agent comprising one or more triazolopyrimidine(s)and/or triazolopyridines described herein, or a tautomer(s) orstereoisomer(s) thereof, or pharmaceutically acceptable salts, solvates,or clathrates of said triazolopyrimidine(s) and/or triazolopyridine(s),said stereoisomer(s), or said tautomer(s), or analogues, derivatives, orprodrugs thereof within the packaging material, along with a secondcomposition comprising a second agent such as, for example, an agentused in the treatment and/or prophylaxis of Alzheimer's disease (e.g.,as described herein), or any prodrugs, co-drugs, metabolites, analogs,homologues, congeners, derivatives, salts and combinations thereof. Insome embodiments, the articles of manufacture may also includeinstructions for using the composition as a prophylactic, therapeutic,or ameliorative treatment for the disease of concern.

EXAMPLES

The following examples are offered to illustrate, but not to limit theclaimed invention.

Example 1 Synthesis of J19

An illustrative, but non-limiting synthesis scheme for J19 is shown inFIG. 3. Reagents and conditions for synthesis Scheme 1 were: (a)2-methoxyethanol, 125° C.; (b) NaNO₂, DCM, acetic acid, r.t; (c) THF,100° C., sealed tube.

6-Chloro-N4-(4-chloro-2-methylphenyl)-2-methylpyrimidine-4,5-diamine (3)

A 250 mL round bottom flask equipped with a reflux condenser was chargedwith a mixture of 4,6-dichloro-2-methylpyrimidin-5-amine (2.170 g, 12.2mmol, 1.0 equiv.), 4-chloro-2-methylaniline (1.726 g, 12.2 mmol, 1.0equiv.) and 2-methoxyethanol (100 mL) and the mixture was heated to anoil bath temperature of 125° C. with stirring under nitrogen. After 48hours, TLC (1:1 ethyl acetate:hexane) indicated completion of thereaction. The mixture was concentrated under reduced pressure to leave aviscous oil. This was dissolved in ethyl acetate (approximately 30 mL)and the desired product precipitated upon the addition of hexane withstirring (approximately 100 mL). The mixture was allowed to sit at 4° C.overnight and the solid was collected by filtration, washed with hexaneand dried under vacuum to give the product as a tan solid (2.4 g, 69%).¹H NMR (CDCl₃. 300 MHz): δ 7.81 (d, J=9.3 Hz, 1H), 7.21-7.19 (m, 2H),6.79 (bs, 1H), 3.29 (bs, 2H), 2.49 (s, 3H), 2.29 (s, 3H). LC/MS: 283.3(M+1).

7-Chloro-3-(4-chloro-2-methylphenyl)-5-methyl-3H-[1,2,3]triazolo[4,5-d]pyrimidine(4)

Sodium nitrite (0.289 g, 4.2 mmol, 1.1 equiv.) was added to a vigorouslystirring mixture of6-chloro-N4-(4-chloro-2-methylphenyl)-2-methylpyrimidine-4,5-diamine(1.060 g, 3.7 mmol, 1.0 equiv.) in dichloromethane (15 mL) and aceticacid (15 mL) at room temperature. After 45 minutes, TLC indicatedcomplete disappearance of the starting material (1:3 ethylacetate:hexane). The mixture was transferred to a separatory funnel and50 mL of water was added. The dichloromethane layer was removed andwashed with water, brine and dried over magnesium sulfate. The organiclayer was then concentrated to dryness to leave a tan solid which wasused directly without further purification (1.10 g, 100%). 1H NMR(CDCl3, 300 MHz): 7.46-7.35 (m, 3H), 2.85 (s, 3H), 2.19 (s, 3H). LC/MS:294.2 (M)⁺

3-(4-Chloro-2-methylphenyl)-N-(1-methoxybutan-2-yl)-5-methyl-3H-[1,2,3]triazolo[4,5-d]pyrimidin-7-amine(6)

A mixture of7-chloro-3-(4-chloro-2-methylphenyl)-5-methyl-3H-[1,2,3]triazolo[4,5-d]pyrimidine(1.0 g, 3.4 mmol, 1.0 equiv.) and 1-methoxybutan-2-amine (0.89 g, 8.9mmol, 2.5 equiv.) in THF (15 mL) was heated in a sealed reaction vesselto an oil bath temperature of 100° C. with stirring. After 3 hours, TLC(20% ethyl acetate in hexanes) indicated complete disappearance ofstarting material and formation of a single product. The mixture wasallowed to cool to room temperature. The mixture was concentrated todryness and purified by flash chromatography over silica gel (0 to 100%ethyl acetate/hexane gradient) to give the title product as a whitesolid after drying. (0.47 g, 60.8%). ¹H NMR (CD3OD, 300 MHz): δ7.54-7.53(m, 1H), 7.46-7.38 (m, 2H), 4.69-4.59 (m, 1H), 3.63-3.50 (m, 2H), 3.40(s, 3H), 2.49 (s, 3H), 2.12 (s, 3H), 1.90-1.62 (m, 2H), 1.02 (t, 7.5 Hz,3H). ¹³C-NMR (75 MHz, CD3OD) δ 9.89, 16.87, 24.45, 24.99, 25.09, 51.60,55.06, 58.17, 74.04, 74.53, 126.96, 128.98, 131.00, 132.83, 135.97,137.80, 150.12, 155.01, 168.03. LC-MS (m/z): 360 [M]+, 362 [M+2]+

Example 2 Evaluation of J03 and Analogs

As described below, in vitro studies were performed using SHSY-5Y cells.These cells are a useful neurological model that differentiate intocells with morphological and biochemical characteristics of matureneurons including mature isoforms of tau.

Animal experiments were performed using J20 mice. The J20 mouse is amodel of for Alzheimer's disease. This model overexpresses human APPwith two mutations linked to familial Alzheimer's disease (the APPKM670/671NL (Swedish) and SAPP V717F (Indiana) mutations).

Initial J03 Studies.

As shown in FIG. 4, panels A-C, J03 reduces tau and phospho-tau (p-tau)increases induced by CRF. In an initial study, SHSY-5Y cells werecultured without serum to induce differentiation and increase tauexpression. CRF was added to the cultures at increasing concentrationsand, as shown in FIG. 4, panels B and C, both tau (panel B) andphospho-tau (panel C) increased with CRF in a dose-dependent manner. Theincreases were proportional and both were about 50%. J03 (FIG. 4A)reduced the CRF-induced tau increases more than the p-tau increases. Theincrease in p-tau with CRF was expected, but the increase in tau lesspredictable as CRF reduces neuronal differentiation (see Chen et al.(2004) Proc. Natl. Acad. Sci. USA, 101(44): 15782-15787), however, itcan increase tau accumulation.

A standard pharmacokinetic study of J03 was performed in which J03 (10mk delivered in a 5 mg/ml DMSO stock, 50 ml) was injected subcutaneously(SQ) into J20 mice. As shown in FIG. 5, J03 brain levels were low andthe brain/plasma ratio was 1:2, but levels stayed up for hours,resulting in good exposure of brain tissue, even from a singleinjection. After oral delivery by feeding (not gavage) also at 10 mg/kg,brain levels were slightly higher at the peak (open grey box), and thebrain:plasma ratio was close to 1:1.

In pilot study #1, J20 mice housed singly were treated by SQ injectionof J02 in PEG/b-MCD at 10 mkd for 12 days. NOR analysis of object memory(N=5/group) was performed. The results are illustrated in FIG. 6, panelsA-F. As shown there was a slight improvement in behavior in singlehoused J03-treated J20 mice (panel A). There was no difference in Aβ1-40(panel B), but a decrease in Aβ1-42 (panel C). The Aβ1-40/1-42 ratio wassignificantly increased (panel D). SAPP was very slightly increased(panel E), as was the sAPP/Aβ1-42 ratio (panel F).

Total tau was decreased by J03 (FIG. 7, top left panel), but largely dueto one mouse. P-tau was significantly lower (FIG. 7, top right panel),and the ratio was slightly higher, but with great individual variation(FIG. 7, bottom panel).

Based on these promising results, the study was repeated (pilot study#2) with oral delivery and a longer duration.

As illustrated in FIG. 8, J03 treated mice performed well in both novellocation (left) and novel object (right) assays.

As shown in FIG. 9, sAPPα increased slightly (top left), sAPPβ wasunchanged (top right), and the sAPPα/sAPPβ ratio (bottom panel) wasslightly increased.

Both Aβ1-40 (FIG. 10, top left) and Aβ1-42 (FIG. 10, top right) wereunchanged with no significant trend to increase. The Aβ1-40/Aβ1-42 ratio(FIG. 10, bottom) was unchanged.

FIG. 11 illustrates the effects of J03 on Aβ1-42 in pilot studies #1 and#2). All mice in the pilot study #2 were older than the mice used in J03pilot study #1. This was deliberate (as was the choice of younger micein the first study) as it appeared in the first study the greatestdifference in Aβ1-42 was in the oldest J03-treated mouse. When theresults are graphed together (with adjustment for dilution factor in theassay), the oldest vehicle-treated mouse in J03 pilot study #1 appearsto be a slightly high outlier. Nonetheless, the curves here suggestthere is some Aβ-decreasing effect if treatment is started in youngermice, but it is lost if treatment is started after Aβ amplification isunderway, as it was in the oldest mice in J03 pilot study #2.

FIG. 12 shows the effect of J03 on p-tau in pilot study #2. Not only wasp-tau decreased again here as it was in pilot study #1, the decreasereached statistical significance.

FIG. 13, panels A-F, shows the effect of J03 on p-tau and memory. Evenvehicle-treated J20 mice showed some novel object preference in themodified protocol, with the erro bar extended to 8 (panel A). There wasgood correlation to p-tau levels (panel B). Mice that scored above 8 hadsignificantly lower p-tau levels (panel C). Novel location preferencewas even clearer for J03-treated J20 mice (panel D), and also showedgood correlation to p-tau levels (panel E), and mice that scored over 0,therefore showing some novel location preference, had significantlylower p-tau (panel F).

To obtain an early look at plasma and brain J03 levels after oral dosingin formulation, two additional mice were dosed on the first day of thestudy and euthanized two hours later. In these mice brain levels werehigher than expected, at 170 ng/g, (FIG. 9, left panel) but with greatvariation between the two mice. At the end of the study brain levelswere in the expected range of about 35 ng/g (FIG. 9, right panel), againwith great variation.

J04 Studies.

J04 (see, FIG. 2) was designed as an analog with replacement of thetriazolopyridine with the triazolopyrimidine ring.

In a primary screen in SH-Sy5Y cells, J04 did not lower tau in theprimary screen, but did lower p-tau, although not significantly (see,FIG. 15). However, J04 did lower p-tau to the same level as J03 andsignificant may not have been reached due to low N #.

Brain levels after SQ injection or oral delivery (FIG. 16, left) weresimilar at −55 ng/g, although clearance after injection was slower.Clearance after oral delivery is shown in FIG. 16, right.

J17 Studies.

In compound J17 (see FIG. 2), two substituents (see circled substituentsin FIG. 17) were reversed.

J17 was observed to significantly lower tau (FIG. 18, left), and p-tau(FIG. 18, right) in SH-Sy5Y cells and the effect was greater than thatmeasured for J03.

As shown in FIG. 19, after exposure to CRF, sAPPα (top left) was thesame as control for J17, but lower for J03 in this experiment. Tau wasdecreased for both J03 and J17 (top right). While the J17 decrease looksthe same as J03, the value for J17 just missed significance. P-tau wasdecreased for both J03 and J17 (bottom left). The N number of 3 limitedthe ability to reach statistical significance.

FIG. 20 illustrates the effect of J03 and J17 on sAPPα, tau, and p-tauwith increasing concentrations of CRF. The decrease in sAPPα with J17,or rather, a lack of increase, was seen without CRF and at 50 nM (topleft), but not at other concentrations. The decrease in tau was seenwithout CRF and at 50 and 100 nM (top middle). The decrease in p-tau wasseen at all concentrations of CRF tested, excluding 0 (bottom left).

FIG. 21 illustrates in vivo pharmacokinetics of J17. After subcutaneousinjection (SQ Inj) of 10 mg/kg, brain levels were low (˜400 ng/g)relative to plasma levels (˜4000 ng/m) (left). After oral delivery atthe same dose, brain levels were only ˜50 ng/g and plasma levels wereonce again almost 10-fold higher.

In a J17 pilot study (#1 pilot study) male and female J20 mice werehoused singly and treated with J17 at 10 mkd by oral delivery for 28days. Male and female J20 mice were housed singly and treated with J17at 10 mkd by oral delivery for 28 days.

The results for males and females showed some differences and arepresented together and separately in FIG. 22. As shown therein, both J03and J17 lowered activity, but not significantly due to individualvariation (top panel). Overall, J03 increased novel object preferencemore than 17, but only the increase with J17 was significant as therewas less variation (bottom left). Males (n=6 per group) showed a patternsimilar to all the mice (bottom middle panel), with the improvement inmemory being less and more similar between J03 and J17. While thegreatest increase in NOP was in a female with J03 (bottom right panel),it was only one female (there was also only one female in the NTggroup). There were only two females in the J03 and J17 groups.

FIG. 23 shows the effect of J03 and J17 on sAPPα, sAPPβ, and the ratiosAPPα/sAPPβ. The sAPPα results were similar for both genders and showedan increase with J03 but not J17 (top panel). sAPPβ results were alsosimilar. There was an decrease with J17 only (one high outlier that waseliminated from the data) (middle panel). There was only a slightincrease in the ratio for males with J03, and it was greater with J17,but the two females showed an even greater increase in the ratio, butwithout statistical significance (bottom panel).

When data from all mice are presented (FIG. 24, top panel), there are nosignificant differences in Aβ1-42, but when the low outlier (10-foldlower) from one sibling pair was eliminated, there is a significantreduction in Aβ1-42 in male mice (FIG. 24, bottom panel). Even with lowoutliers eliminated, there was no significant difference in theAβ1-40/Aβ1-42 ratio (FIG. 25).

FIG. 26 A 1-42. When data from all mice are presented (top), there areno significant differences in Aβ1-42 (FIG. 26, top), but when the lowoutlier (10-fold lower) from one sibling pair was eliminated, there wasa significant reduction in Aβ1-42 in male mice (FIG. 26, bottom).

Tau (FIG. 27, top panel) was increased in males, but an importantreadout for this series of compounds, p-tau, was slightly lower in malemice (FIG. 27 middle panel) and the p-tau/tau ratio (FIG. 27,bottompanel) was lower still and just missed significance forJ17-treated males.

J19 Studies.

J19 (see FIG. 2) is an analog of J03 similar to J17 and is part of thetriazolopyrimidine series described herein.

FIG. 28, panels A-D, illustrates the effect of J03 and J19 onCRF-induced tau and p-tau alterations. J03 significantly decreased tauboth in the absence of CRF and in the presence of 100 nM CRF (panel A).J19 also showed a trend to decrease tau. Both J03 and J19 significantlyreduced p-tau in the presence and absence of CRF (panel B). Thep-tau/tau ratios were highly significantly decreased by J03 and J19 atboth 50 and 100 nM CRF, and were decreased in the absence of CRF (panelC). There was a trend for sAPPα to be higher with J03 and J19 thatreached significance for J03 at 100 nM CRF (panel D).

J19 decreased tau and both J03 and J19 decreased p-tau (FIG. 29, leftpanel). Therefore, while the p-tau/tau ratio looks unchanged for J19FIG. 29, right panel). This is due to the tau decrease and not due to alack of effect.

FIG. 30 illustrates the in vivo pharmacokinetics of J19. After SQinjection of J19 at 10 mkd, brain levels were ˜190 ng/g at the peak 4hours after injection. Levels remained relatively high until 8 hours(left panel). With oral delivery (right panel), levels were much lower,but again remained detectable from 1 to 6 hours.

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It is understood that the examples and embodiments described herein arefor illustrative purposes only and that various modifications or changesin light thereof will be suggested to persons skilled in the art and areto be included within the spirit and purview of this application andscope of the appended claims. All publications, patents, and patentapplications cited herein are hereby incorporated by reference in theirentirety for all purposes.

1. A compound according to the formula:

or a pharmaceutically acceptable salt, stereoisomer, or tautomerthereof, wherein: R⁰ is present or absent, and when present is NH orNCH₃;

and R¹ is

or is selected from the group consisting of: pyridine; pyrimidine;naphthalene; quinolone; isoquinoline; cinnoline; phenyl; oxazole; furan;pyran; isoxazole; thiazole; thiophene; pyrrole; pyrrolidine; pyrazole;imidazole;

wherein R⁴, R⁵, and R⁶ are independently selected from the groupconsisting of H, OH, halogen, methyl, OCH₃, OCF₃, OCHF₂, N(CH₃)₂, ethyl,propyl, butyl, NH-alkyl, O-alkyl, and SO₂CH₃; and

wherein R⁷ and R⁸ are independently H, CH₃, OCH₃, and halogen; or R¹ is

and R² is

or is selected from pyridine; pyrimidine; naphthalene; quinolone;isoquinoline; cinnoline; phenyl; oxazole; furan; pyran; isoxazole;thiazole; thiophene; pyrrole; pyrrolidine; pyrazole; imidazole;

wherein R⁴ and R⁶ are independently selected from the group consistingof H, OH, halogen, methyl, OCH₃, OCF₃, OCHF₂, N(CH₃)₂, ethyl, propyl,butyl, NH-alkyl, O-alkyl, and SO₂CH₃, and R⁵ is H, OCH₃ or halogen; and

wherein R⁷ and R⁸ are independently H, CH₃, OCH₃, and halogen; R³ isselected from the group consisting of H, CH₃, ethyl, propyl, butyl, CF₃,NH₂, halogen, and CH₂O where R is H, alkyl, or aryl; and said compoundis not J03, J04, J05, J08, and J17.
 2. The compound of claim 1, or apharmaceutically acceptable salt, stereoisomer, or tautomer thereof,wherein R¹ is


3. The compound of claim 2, or a pharmaceutically acceptable salt,stereoisomer, or tautomer thereof, wherein R² is

where R⁴ and R⁶ are independently selected from the group consisting ofH, OH, halogen, methyl, OCH₃, OCF₃, OCHF₂, N(CH₃)₂, ethyl, propyl,butyl, NH-alkyl, O-alkyl, and SO₂CH₃ and R⁵ is H, OCH₃, or halogen. 4.The compound of claim 1, or a pharmaceutically acceptable salt,stereoisomer, or tautomer thereof, wherein R² is


5. The compound of claim 4, or a pharmaceutically acceptable salt,stereoisomer, or tautomer thereof, wherein R¹ is

where R⁴, R⁵, and R⁶ are independently selected from the groupconsisting of H, OH, halogen, methyl, OCH₃, OCF₃, OCHF₂, N(CH₃)₂, ethyl,propyl, butyl, NH-alkyl, O-alkyl, and SO₂CH₃.
 6. (canceled)
 7. Thecompound of claim 3, or a pharmaceutically acceptable salt,stereoisomer, or tautomer thereof, wherein R⁵ is OCH₃ or halogen.
 8. Thecompound of claim 7 or a pharmaceutically acceptable salt, stereoisomer,or tautomer thereof, wherein R⁴ is CH₃ or OCH₃ and R⁶ is H or CH₃. 9-16.(canceled)
 17. The compound of claim 5 or a pharmaceutically acceptablesalt, stereoisomer, or tautomer thereof, R⁵ is F, Cl, CH₃ or OCH₃. 18.The compound of claim 17 or a pharmaceutically acceptable salt,stereoisomer, or tautomer thereof, wherein R⁴ is CH₃, halogen, or OCH₃and R⁶ is H or CH₃. 19-24. (canceled)
 25. The compound of claim 2, or apharmaceutically acceptable salt, stereoisomer, or tautomer thereof,wherein R² is

where R⁷ and R⁸ are independently H, CH₃, OCH₃, and halogen.
 26. Thecompound of claim 4, or a pharmaceutically acceptable salt,stereoisomer, or tautomer thereof, wherein R¹ is

where R⁷ and R⁸ are independently H, CH₃, OCH₃, and halogen. 27.(canceled)
 28. The compound of claim 2, or a pharmaceutically acceptablesalt, stereoisomer, or tautomer thereof, wherein R² is


29. The compound of claim 4, or a pharmaceutically acceptable salt,stereoisomer, or tautomer thereof, wherein R¹ is

30-31. (canceled)
 32. The compound of claim 1, or a pharmaceuticallyacceptable salt, stereoisomer, or tautomer thereof, wherein R⁰ is NH.33. (canceled)
 34. The compound of claim 1, or a pharmaceuticallyacceptable salt, stereoisomer, or tautomer thereof, wherein saidcompound is a compound according to a formula selected from the groupconsisting of

35-38. (canceled)
 39. A pharmaceutical formulation comprising; acompound of claim 1, or a pharmaceutically acceptable salt,stereoisomer, or tautomer thereof; and a pharmaceutically acceptablecarrier or excipient. 40-41. (canceled)
 42. A method of decreasing p-tauin a mammal or inhibiting or preventing an increase in p-tau in amammal, said method comprising: administering to said mammal aneffective amount of one or more compounds of claim 1; and/oradministering to said mammal an effective amount of a compound selectedfrom the group consisting of J03, J04, J05, J08, and J17, or apharmaceutically acceptable salt or solvate thereof.
 43. A method ofpromoting the processing of amyloid precursor protein (APP) by thenon-amyloidogenic pathway in a mammal, said method comprising:administering to said mammal an effective amount of one or morecompounds of claim 1; and/or administering to said mammal an effectiveamount of a compound selected from the group consisting of J03, J04,J05, J08, and J17, or a pharmaceutically acceptable salt or solvatethereof.
 44. (canceled)
 45. A method of preventing or delaying the onsetof a pre-Alzheimer's condition and/or cognitive dysfunction, and/orameliorating one or more symptoms of a pre-Alzheimer's condition and/orcognitive dysfunction, or preventing or delaying the progression of apre-Alzheimer's condition or cognitive dysfunction to Alzheimer'sdisease in a mammal, said method comprising: administering to saidmammal an effective amount of one or more compounds of claim 1; and/oradministering to said mammal an effective amount of a compound selectedfrom the group consisting of J03, J04, J05, J08, and J17, or apharmaceutically acceptable salt or solvate thereof.
 46. A method ofameliorating one or more symptoms of Alzheimer's disease, and/orreversing Alzheimer's disease, and/or reducing the rate of progressionof Alzheimer's disease in a mammal, said method comprising:administering to said mammal an effective amount of one or morecompounds of claim 1; and/or administering to said mammal an effectiveamount of a compound selected from the group consisting of J03, J04,J05, J08, and J17, or a pharmaceutically acceptable salt or solvatethereof. 47-81. (canceled)